Comparison of Umbilical Cord Blood vs Personalized Treatment for Improving Autistic Spectrum Disorder
Phase: PHASE1
Conditions: Autistic Spectrum Disorder
Sponsor: Fundatia Bio-Forum

Protocol Version 1: 10 January 2019. Corrected 26.06.19; 17.06.21, Revised 14 October 2021             
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Bio-Forum Foundation
Clinical Study Protocol No: B-F011  
 
                           
 
                                                
 
   Protocol Date: 14 
 
 October
 
  2021
 
 
Protocol Cover Page
Identifiers:
NCT04007224     Unique Protocol ID: CORDUS
Protocol Title:
A pilot crossover study to investigate the safety and efficacy of
administering  intravenous  autologous  cord  blood  versus
personalized  noninvasive  treatments
 to 
children  with
developmental disorders of the autistic spectrum.
Brief Title:
Umbilical Cord Blood vs Personalized Treatments for Improving 
Autistic Disorder
Protocol Number:
Bio-Forum Foundation 011
(B-F011)
Protocol Date:
14 October 2021
Study Phase:
I-II
Principal Investigator:
Felician Stancioiu, MD, Ruxandra Caisan, MD, Lorena Ulea, MD,
Investigators:
Radu Dumitrescu MD, Raluca Bogdan, MD, Mihaela Agache,  
MD, Laura Ion, MD
Research Team: 
Felician Stancioiu, MD, Livia Sima, PhD, Bogdan Ivanescu, MD  
Protocol Author(s): 
Felician Stancioiu, MD
Confidentiality Agreement
This document is a confidential communication of The Bio-Forum Foundation and can be read and used
only with the implicit agreement by the recipient that no unpublished information contained within will
be published or disclosed without prior written approval.
SPONSOR:    
Bio-Forum Foundation
 
Blv D Cantemir 2B, 
Bl P1, Sc 1 Apt 4
Bucharest, Romania
INVESTIGATOR:        
Felician Stancioiu, MD
Spitalul Angiomedica
Str Cosminului Nr 13
Bucuresti, Romania

Protocol Version 1: 10 January 2019. Corrected 26.06.19; 17.06.21, Revised 14 October 2021             
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Clinical Study Protocol No: B-F011  
 
                           
 
                                                
 
   Protocol Date: 14 
 
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  2021
 
 
TABLE OF CONTENTS
PAGE
              LIST OF ABBREVIATIONS AND DEFINITIONS
4
STUDY CONTACT LIST
5
               
PROTOCOL SYNOPSIS
6
1            INTRODUCTION
8
1.1
Abstract
8
1.2
Background
8
1.3
Study Rationale
9
2
STUDY OBJECTIVES
10
3
STUDY PLAN AND PROCEDURES
11
3.1
Study design 
11
3.1.1
Scheduled clinic visits
11
3.1.2
Visits and assessments
11
3.2
Study population
12
3.2.1
Inclusion criteria
12
3.2.2
Exclusion criteria
12
3.2.3
Justification for inclusion and exclusion criteria 
13
3.2.4
Criteria for discontinuation
13
3.3
 Investigational Products and Treatments
13
3.3.1
Treatment Schedule
13
3.3.2
Randomization
13
3.3.3
Identity of Study  Treatments
14
3.3.4
Storage and Accountability
14
3.3.5
Allowed medication
14
3.4.1
Compliance
14
4
STUDY MEASUREMENTS AND ENDPOINTS
14
4.1 Primary Safety Endpoints
14
4.2 Secondary Safety Endpoint
14
4.3 Measurements at each visit
15
4.4 Specific detail on measurements
16
4.4.1
Adverse Events
16
4.4.2
Laboratory Investigations
16
 
                                                              
5
STATISTICAL METHODS 
18
5.1 Determination of sample size
18
5.2 Statistical analysis
18
5.3 Changes to the protocol
18

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Clinical Study Protocol No: B-F011  
 
                           
 
                                                
 
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6
ETHICS
19
6.1 Ethics review
19
6.2 Ethical conduct of the study
19
6.3 Subject information and consent
19
6.4 Subject data protection 
19
7
DATA QUALITY ASSURANCE
19
8
STUDY TIMETABLE AND TERMINATION
20
9
REFERENCES
20
10
APPENDICES
1.1   Appendix A - 
Declaration of Helsinki
22
1.2   Appendix 1 - Sample written informed consent          
25
1.3 . Annex 1 – Costs
31
1.4   Annex 2 – CAST  questionnaire
32
1.5 Annex 3- M-CHAT questionnaire
34
1.6 Annex 4 – Q-CHAT  questionnaire 
36
1.7  Annex 5 - UCB thawing protocol (Duke/Rubinstein protocols by Vita 34/)
40
1.8.Annex 6 – UCB infusion 
44

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Clinical Study Protocol No: B-F011  
 
                           
 
                                                
 
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LIST OF ABBREVIATIONS AND DEFINITIONS
The following abbreviations and specialist terms are used in this study protocol:
Abbreviation 
Explanation
AE
Adverse Event
ALT
Alanine aminotransferase
ASD
Autistic Spectrum Disorder
AST
Aspartate aminotransferase
b.i.d
bis in die = twice daily
CD45+
count of CD45 expressing cells
CNS
Central Nervous System
CRF
Case Report Form
ECG
Electrocardiogram
FC
FlowCytometry
GCP
Good Clinical Practice
GGT
gamma glutamyl transpeptidase
Hb
Haemoglobin
Hct
Haematocrit
HDL-C
High Density Lipoprotein –cholesterol
HLA
Human Leukocyte Antigen 
ICH
International Conference of Harmonisation
IEC
Institutional Ethics Committee
IP
Inflammatory Panel
IRB
Institutional Review Board
LCMS
Liquid chromatography linked mass spectrometry
LDH
Lactate dehydrogenase
LDL-C
Low Density Lipoprotein -cholesterol
MB
Microbiology (testing)
MCH
Mean corpuscular haemoglobin
MCHC
Mean corpuscular haemoglobin concentration
MRI
Magnetic Resonance Imaging
ml
milliliter
PQQ
Pironoquinolinquinone
RDW
Red cell distribution width
RBC
Red Blood Cells
SAE
Serious Adverse Event
SEM
Standard error of the mean
T3
Triiodothyronine
T4
Thyroxine
TSH
Thyroid-stimulating hormone
UCB
Umbilical Cord Blood
WBC
White blood cells
 

Protocol Version 1: 10 January 2019. Corrected 26.06.19; 17.06.21, Revised 14 October 2021             
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Clinical Study Protocol No: B-F011  
 
                           
 
                                                
 
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STUDY CONTACT LIST
For questions regarding the conduct of this study, please contact:
Project Leader:
Felician Stancioiu, MD
E-mail:
f elicians@bio-forum.net
 
 
Mobile:
0040727500402
Study Coordinator:
Bogdan Ivanescu, MD
 
E-mail:
bogdan.ivanescu@doctornit.ro
Telephone:
0722504540
 
 
For reporting a serious adverse event, please contact:
 Principal Investigator:
Felician Stancioiu, MD
E-mail:
f elicians@bio-forum.net
 
 
Mobile:
0040727500402
For questions regarding data quality assurance, please contact:
Independent Study Monitor:
Ioana Scantei, RN
Mobile:
0763640163

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Clinical Study Protocol No: B-F011  
 
                           
 
                                                
 
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PROTOCOL SYNOPSIS
TITLE
A pilot crossover study to investigate the safety and efficacy of 
administering intravenous autologous cord blood versus personalized 
noninvasive treatments to children with developmental disorders of the 
autistic spectrum
Investigational site:
Spitalul Angiomedica, Str Cosminului Nr 13, Bucharest, Romania
Investigators:
Dr Felician Stancioiu, Dr Ruxandra Caisan, Dr Lorena Ulea, Dr Raluca 
Bogdan, Dr Radu Dumitrescu, Dr Laura Ion, Dr Mihaela Agache
Sponsor:
Bio-Forum Foundation, Bucharest, Romania
Representatives:
Dr Felician Stancioiu
Study number:
B-F001
Final Protocol:
June 2019
Clean File:
June 2019
Ethics Approval:
January 2019
Statistical analysis:
 
 
Clinical Phase:
 
Study Report:
 
OBJECTIVES:
To compare the safety and efficacy of administering intravenous 
autologous cord blood versus personalized noninvasive treatments to 
children with developmental disorders of the autistic spectrum
STUDY DESIGN:
An open-label, pilot crossover study over a 12-month period.  Participants
will be screened and randomized at Visit 0 and at visit V1 will be receiving
either intravenous umbilical cord blood or noninvasive treatment. At V2, at
2 months after V1, the other treatment is administered and subsequently
the child will be evaluated by questionnaires and child therapist.   Parents
will report adverse events and complete the M-CHAT, Q-CHAT and CAST
questionnaires.  Modifications  on  these  questionnaires  will  be  analized
statistically as a quantitative measure of child’s improvement.
SUBJECTS:
Inclusion criteria
Males and females between 3 and 7 years of age:

parents informed of the nature of the study and will give written
informed consent,

have body weights between 10-30 kg,

have no clinically significant abnormal values on hemoleucogram
during screening,

be on no regular medical treatment

have no known allergies to treatments
Exclusion criteria

Any  genetic  or  metabolic  disease  or  condition  which  might
compromise  the  haematopoietic,  renal,  endocrine,  pulmonary,
central  nervous,  cardiovascular,  immunological,  dermatological,
gastrointestinal or any other body system.

History of allergic conditions – asthma, urticaria, eczema.

History of autoimmune disorders   

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
Intake of any medication within 14 days before start of the study

Presence of significant abnormal hemoleucogram results during
screening.
TREATMENT TO 
BE EVALUATED
Active Treatment A:  Autologous umbilical cord blood administered 
intravenously as single treatment .
Active Treatment B:  Personalized daily treatment following evaluation 
with a combination of intranasal Oxytocin; Curcumin; Lecithin, Piracetam, 
Tecfidera; PQQ; Magnesium citrate and vitamin B6
DURATION OF 
STUDY
12 months 
ENDPOINTS:
EFFICACY
       1.    Behavioral improvement with the validated scales for autistic 
spectrum disorders
       2.    Examination by child psychologist
SAFETY 
 
  
1.
Adverse  events, especially frequency and  intensity of agitation
episodes
2.
Modification of vital signs:  blood pressure, heart rate, respiratory 
rate, body temperature.
3.
Modification of laboratory tests: Haematology: RBC, MCHC, MCH, 
MCV, RDW, Hct, Hb, WBC Differential count –lymphocytes 
monocytes, neutrophils, eosinophils, basophils; CD45+; blood 
cultures
Biochemistry: Serum - Bilirubin (total) AST, ALT,  LDH, glucose, 
urea, creatinine, sodium, potassium, chloride, calcium., 
MB microbiology; IP – Inflammatory panel
  
 
SAMPLE SIZE:
at least 25 patients, optimum 40 patients enrolled
STATISTICAL 
ANALYSIS:
Summary statistics will describe the safety variables.  Significance testing
will be done using analysis of variance for continuous and chi-square test
for categorical variables.  (α = 0,05)

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1.   INTRODUCTION
1.1 Abstract
1.
The treatment of children with autistic spectrum disease (ASD) represents a serious challenge
for both the patients and their families, and also the medical staff involved.  
Recently (Dawson G, 2017) prof Geraldine Dawson from Duke University showed that 
the use of stem cells from autologous umbilical cord blood (UCB) is a safe and efficacious 
therapy, opening a new and exciting avenue for therapeutic redress in children with ASD. This 
new therapeutic venue has yielded more dramatic improvements than any previous treatment 
including intranasal oxytocin. 
We know that ASD has multiple determinants - genetic, environmental, neurological, and 
immunological (Young 2016) – and the pathological processes involved in ASD are various and 
complex. ASD is after all a spectrum of diseases involving brain development, not just a well-
defined condition, and the implication is that the one-molecule-fits-all type of treatment will not 
be likely to address the multitude of aspects and deficiencies which are present in individuals 
with ASD. UCB itself contains a plethora of molecules which can modulate the activities of 
neurons and the supporting microglia, so comparing it with just one therapeutic substance would 
not be a good strategy.
With this in mind, we propose that ASD treatment should be very much personalized after a 
careful evaluation of the patient, and a noninvasive treatment option should include two or more 
substances from the following: oxytocin, curcumin, magnesium citrate with vitamin B6, 
pironoquinolinquinone (PQQ), lecithin, piracetam, and optionally dimethylfumarate, a 
medication known to be both neuroprotective and immunomodulatory.
Below we propose a pilot clinical study which aims to compare the safety and efficacy of 
UCB administration versus a less invasive and costly therapeutic option which consists of a 
patient-specific combination of medications and supplements. The study will be open-label, 
crossover type, meaning that all children will get sequentially both treatments after a 2-week 
washout interval. This study design has the advantage of eliminating the ethically questionable 
administration of a placebo. 
1.2 Background
There is ample information in the medical literature on ASD, due to the increasing incidence 
of the condition, impact on families and society and the healthcare costs associated with it.
As it affects the neuropsychological status of the developing child, it is classified and studied 
as a psychiatric ailment, and as such DSM-5, characterizes ASD behavior by the following 
traits:
- difficulties in social reciprocity and communication,
- unusually narrow interests,
- repetitive behaviours and speech,
- insistence on sameness, and
- idiosyncratic sensory responses

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We also know that genetic, environmental, neurological, and immunological factors 
contribute to its etiology and among these we mention: 
- increased rate of amygdala growth (Nordahl, 2012)
- atypical pruning of synapses (Geschwind 2007)
- decreased connectivity between ventromedial prefrontal cortex, which is implicated in 
emotion and social communication and the middle temporal gyrus/superior temporal sulcus 
region, which is implicated in face expression processing involved in social behaviour; also 
the precuneus/superior parietal lobule region with reduced functional connectivity, which is 
implicated in spatial functions including of oneself, and of the spatial environment (Xydakis 
2015)
- neuron density in the prefrontal cortex was shown to play a significant role (Courchesne 
2011)
- neuroinflammation in the anterior areas of the cortex (Pardo, 2005;
- increased head size in ASD children correlated with a history of allergic/immune disorders, 
(Sacco, 2007, Nordahl, 2011)
- increased titers of maternal autoimmune antibodies (Croen 2008, Braunschweig 2012)
Considering all the above, the consensus is that he prevailing pathological modification in 
children with ASD is the presence of a persistent pro-inflammatory status in the central 
nervous system (CNS) of the patient (Young 2016). 
There are not many treatment options which are known to be effective in treating ASD. 
Secretin, oxytocin (Parker 2017), vaccines were among the treatments studied, but the results
of the respective clinical studies were not at all encouraging. 
1.2
Study Rationale
        
       Even though it is a recent development, UCB has had the best results so far in treating 
children with ASD and its results recommend it for treatment to any child with ASD and who
had UCB collected at birth. Two important questions arise: 
- what other treatment can be recommended to the ASD children without UCB collected at 
birth, and with what comparative efficacy, and
- being a one-time intervention, can we improve/add to UCB infusion other substances which
have a very good safety profile (well tolerated) and at least theoretically have a benefit in 
improving neuronal function?
To begin to answer these questions we have devised an open-label clinical study with a 
crossover design (all children receive both treatments, albeit in a different sequence). 
One important novelty with our study is that the treatment to which UCB is compared is not 
the same to all children, but is individually tailored to address specifically the most important
problems the child has (inflammation, anxiety/social avoidance, myelination, global delay, 
etc)
Based on the results of the testing with the M-CHAT-R, Q-CHAT and CAST questionnaires 
and the evaluation from child psychologist, a recommendation for treatment will be made. 
The noninvasive treatment consists of a combination of at least two substances from the 

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following: Oxytocin 10 UI/day intranasally, Magnesium citrate and vitamin B6 ½ cp qd, 
pironoquinolinquinone (PQQ) 20 mg qd, Lecithin 1000 mg qd, Curcumin 500 mg qd.
The specific treatment components will be recommended and administered according to the 
criteria below, after performing all the tests (imagistics, labs, questionnaires) which can 
suggest a deficiency on one or more of the following directions:
- inflammatory problem (IP markers on labs, white matter changes, diffuse or no specific 
modification on MRI)
- myelination (MRI changes),
- anxiety/social avoidance (questionnaires),
- severe developmental delays in cognition, logic, verbalization
1. Curcumin 250 mg bid will be included in the child’s regimen if the imagistics (MRI/CT 
scan) and/or laboratory values (IP) suggest the presence of an inflammatory status, either in 
the CNS (white matter modifications on the brain scan) or systemic (abnormal values in the 
Inflammatory Panel done from blood);
2. Oxytocin 10 UI/day via intranasal administration will be administered if a social-
communication deficit is clearly shown at evaluation
3. Lecithin 500 mg bid will be administered if brain imaging reveals any 
demyelination/myelination deficits, global cerebral atrophy or regional atrophy  
4. Magnesium citrate with vitamin B6 (150 mg/6 mg qd) will be included if 
restlessness/agitation/anxiety is present significantly in the clinical picture
5. Pironoquinolinquinone (PQQ) 20 mg/day will be administered in situations of 
apathic/disinterested behavior, marked introversion, marked developmental delays
6. Optional medication for special situations, with parent’s clear understanding, a few other 
treatment options will be included – ex.
- Tecfidera (dimethyl fumarate) 60 mg bid with food (presence of hypo/demyelination with 
an autoimmune component); immune modulation is involved in ASD as documented in 
Kornberg, 2018
- Boswellia extract 400 mg bid – decreases activation of NfK-beta which is the main 
modulator of both inflammation and neuronal connections
- piracetam (Nootropil) 400 mg bid (presence of severe developmental delays with low motor
coordination and energy levels)
- Bacopa monieri (presence of demyelination in absence of immune over-activation)
- Uridine (inflammation mitigation, mitochondrial function stimulation)
- Hipophae rhamnoides oil (source of omega-7 unsaturated fatty acids for synaptic vesicles)
The decision of which of these components should be given is taken after all tests are done
and the child psychologist/therapist and the study investigator discuss the case and afterwards
discuss the practical issues of administering the recommended treatment with the parents.
2
  STUDY OBJECTIVES
The primary objective of this study is to investigate the comparative efficacy of the UCB 
infusion and the non-invasive treatments for improving the functioning of children with ASD.

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A secondary objective is to determine whether there are significant differences in the safety 
profiles of the two treatments administered
In addition there will be an evaluation of the structural changes in the CNS of the children with 
ASD, done by MRI
3
STUDY PLAN AND PROCEDURES
3.1  Study design 
This is a twelve months long, open-label, crossover study to evaluate the comparative efficacy 
and safety of UCB versus non-invasive treatments in children with ASD. Each participant will 
receive both treatments in a random sequence.
The UCB is autologous, previously harvested at the child’s birth and cryopreserved in liquid 
nitrogen for later use.
The UCB infusion was performed at Angiomedica Hospital, Str Cosminului nr 13 as main site 
and in two additional clinical sites: Medicover Hospital, Str. Pechea Nr 8, and Ponderas 
Academic Hospital, Str Nicolae Caramfil 85A, all in Bucharest, Romania by anesthesiologist and
pediatric physician, based on the Duke protocol.
Prior to UCB infusion, HLA typing of the child will be performed at the National Institute of 
Hematology, as well as the HLA typing of the UCB prior to thawing – for positive identification.
During this time the UCB collection will be kept in liquid nitrogen for a maximum of 72 hours, 
after which thawing and preparing of the UCB for infusion will be performed by Dr Felician 
Stancioiu; the thawing is done in optimal conditions for preserving simultaneously both sterility 
and viability of the cryogenated cells - the Vita 34 or Rubinstein protocols; testing for viability 
will be done with Trypan Blue and flowcytometry. Choosing optimal conditions is based upon 
the minimum number of processing steps necessary (fewest number of centrifugations and 
shortest time of processing, with best recovery of various stem cells) and ensuring sterility.
The non-invasive treatment combination is prescribed by the study investigator after the three 
questionnaires are answered and an evaluation by child therapist/psychologist is peformed 
independently. This treatment is 
3.1.1  Scheduled study visits
 
Recruitment of patients will occur during initial contacts with the study coordinator 
Bogdan Ivanescu who will present the parents with the opportunity to enroll in the study, and 
when general information will be given about the study on timeline, procedures involved, tests, 
costs. 
After agreeing in principle to participate in the study, parents and child will be scheduled for 
Visit 0, at which the patients’ blood is drawn for testing, questionnaires are completed, the 
patient is evaluated by the child therapist and is randomized for receiving treatment A (UCB 
infusion) or B (noninvasive treatment), and the necessary arrangements are made for UCB 
release from the respective banking site.

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Visit 1 is scheduled within 7-14 days from V0, and during this study visit the child will receive 
either treatment A or B. Treatment B (noninvasive) is administered daily for 6 weeks, and is 
followed by 2 weeks with no treatment (washout period)
Visit 2 follows at 8 weeks distance from V1, which includes the 2 week washout period after the 
first treatment is administered. New blood tests are performed, also questionnaires and 
evaluation by child therapist, and the other treatment in sequence is administered.
Visit 3, Visit 4, and Visit 5 are scheduled at 16, 24 and 52 weeks after first treatment was 
administered (V1) and involve the neuropsychological evaluation of the child by the therapist 
and with the 3 questionnaires; brain imaging is recommended at 52 weeks
Visit 0
 V1
V2 
(+ 8 weeks)
V3
(+16 wks)
V4
 (+24 wks)
V5
(+52 wks)
Labs
CBC, LFTs, 
Els, HLA; IP
 
HLA
MB
CBC, LFTs, 
Els, HLA; IP;
MB
Imagis-
tics
MRI/CT
MRI/CT
Neuro/
Psi 
Evalua-
tion
M-CHAT-R;
Q-CHAT; 
CAST;
- Child 
therapist
 x
M-CHAT-R;
Q-CHAT; 
CAST;
- Child 
therapist
M-CHAT-R;
Q-CHAT; 
CAST;
- Child 
therapist
M-CHAT-R;
Q-CHAT; 
CAST;
- Child 
therapist
M-CHAT-R;
Q-CHAT; 
CAST;
- Child 
therapist
Treat-
ment 
started
randomi-
zation
A/B
B/A
X
X
X
CBC – complete blood count with differential; LFTs – liver function tests, Els – electrolytes; IP 
– Inflammatory panel – ex ESR, CRP, serum protein electrophoresis, neopterin, IL-6; HLA – 
Typing of patient HLA- A, HLA-B, HLA-DR; MB - microbiology
3.2   Study population
This clinical study will include children diagnosed with ASD, 3-7 years, and who meet the 
inclusion criteria outlined above.
Recruitment will continue until a minimum of 25 patients have received both treatments. 
3.2.1 Inclusion criteria
To be enrolled, the following criteria have to be fulfilled at Visit 2:
1. Children of either sex between 3 and 7 years. 
2.
Body weight between 15 and 30 kg
3. Diagnosed with ASD
4. No significant diseases or clinically significant abnormal laboratory values during screening
5. On no regular medical treatment  

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6. Parents able to be informed of the nature of the study and willing to give written informed 
consent (Consent must be obtained before any study-related procedures are conducted).
3.2.2
Exclusion criteria
Any of the following is regarded as a criterion for exclusion from the study:
1. Hypersensitivity or allergic reaction to any drugs or supplements.
2. Any disease or condition which might compromise the haematopoietic, renal, endocrine, 
pulmonary, central nervous, cardiovascular, immunological, dermatological, gastrointestinal 
or any other body system.
3. History of allergic conditions – asthma, urticaria, eczema.
4. History or presence of dyspepsia, gastric ulcer or duodenal ulcer.
5. History of autoimmune disorders e.g. systemic lupus erythematosus, haemolytic anemia.
6. Intake of steroid medication within 7 days before start of the study
7. Presence of clinically significant abnormal laboratory results during screening.
8. Participation in a clinical study of any investigational product 1 month prior to visit 1 or 
during the study.
3.2.3
Justification for inclusion and exclusion criteria
The criteria are set to minimize the risk to the patients, to ensure a study population that will 
enable the investigation of the set objectives, and to provide equal opportunity for inclusion. 
3.2.4
Criteria for discontinuation
Subjects may be discontinued from study treatment and assessments at any time, at the discretion
of the investigator. Specific reasons for discontinuing a subject from the study are:
1. Withdrawal of informed consent.
2. Development of exclusion criteria, pregnancy or other safety reasons during the study.
3. Protocol non-compliance.
4. Incorrect enrollment or randomization of the subject.
For subjects withdrawn from the study, the same measurements and assessments should be 
performed as done at Visit 5.  Adverse events should be followed up and questionnaires 
completed by parents.
3.3 Investigational Products and Treatments
3.3.1  Treatment Schedule
Each treatment arm is administered and considered active for a period of maximum 8 weeks; the 
non-invasive treatment is administered for 6 weeks followed by a 2-week “wash-out” period, and
the UCB infusion will be followed by supplement administration after an 8-week interval.
Patient evaluation, registration informed consent and randomization occur at initial visit 0 (V0); 
followed after 14-21 days by V1 (longer for the preparation of UCB infusion), during which the 
first treatment is administered; V2 after another 8 weeks, at which the second treatment is 
administered, and V3, V4, and V5, at 16, 24 and 52 weeks respectively after V1.

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A tabulated summary of the study visits was given above Section 3.1.1
3.3.2
Randomisation
Because it is an open-label study, there is no need for blinding/assigning participant numbers.
Each participant will be randomly assigned to begin the study either with the UCB infusion or 
the non-invasive treatment. 
Eligible patients, who for whatever reason, do not report to the clinic to receive the first 
treatment after having been selected for the study will be referred to as discontinuers in the 
report.  A patient who, for whatever reason, withdraws or is withdrawn from the study after 
having been enrolled, will be classified as a dropout, and identified as such in the relevant Case 
Report Form (CRF).
3.3.3
Identity of study Treatments
After receiving it from the storage bank, UCB will be tested for HLA determinants at the 
National Institute of Hematology laboratory by gene sequencer.
Afterward positive confirmation UCB will be thawed according to the Duke/Rubinstein 
protocols. A vial containing 1.5 ml thawed UCB will be sent for TNC/viability by flowcitometry 
at Victor Babes Laboratory. A label with patients’ name, date is affixed to each UCB bag 
prepared for infusion.
 
3.3.4.
Storage and Accountability
UCB collections will be kept in liquid nitrogen, vapor phase – 170 degrees Celsius -  at all times 
prior to thawing of collection, and there will be periodic recording of this condition. Transport 
recipients (dry shippers come with temperature sensors, which will be photographed, and during 
HLA testing, the collection is kept also in liquid nitrogen and its storage at below  – 170 degrees 
Celsius will also be documented via periodic photography of the measuring device.
The Principal Investigator of the study – Dr Felician Stancioiu – is directly responsible for this 
stage of UCB preparation.
3.3.5.  Allowed medication
Preferably no other medicine should be taken by the subjects. The use of any incidental 
medication  (e.g. mild analgesics, NSAID, etc) must be recorded.
 
3.3.6
Compliance 
Study code:  Bio-Forum Foundation001
Visit no:
Investigator: Dr
                Subject no:
Investigational product: UCB    
Patient name
Keep at room temperature 
Exp Date:
For clinical study use only

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The parents will attest to the use of study medication on a daily basis and the adherence to the 
prescribed treatment will be checked at every clinic visit.
4
STUDY MEASUREMENTS AND ENDPOINTS
In this study the following endpoints will be measured/recorded
1. Efficacy  - difference in the scores on the questionnaires M-CHAT, Q-CHAT and CAST 
before and after each treatment
2. Safety – type, severity, duration, treatment and amelioration of each of the adverse effects 
occurring during respective treatments.
4.1
Primary Safety Endpoints.
  
1. Adverse events  (type and frequency)
2. Physical examination parameters (body mass, height, etc)
3. Vital signs: blood pressure, heart rate, respiratory rate, body temperature 
4. Laboratory tests
HLA testing by genetic sequencing
Haematology: CBC - RBC, HB. MCHC, MCH, MCV, RDW, Hct, Hb, WBC Differential
count –lymphocytes monocytes, neutrophils, eosinophils, basophils, 
flowcytometry - CD34+, TNC, viability %
Microbiology – presence and sensitivity of pathogens
LFTs Serum - Bilirubin (total and conjugated) AST, ALT, GGT, LDH, glucose
IP – serum protein electrophoresis, ESR, CRP, fibrinogen
Els - sodium, potassium, chloride, calcium, magnesium.
4.2
Measurements at each visit.
See section 3.1.1 for the visit schedule.
 
Visit 0: Screening Visit
Patients will be examined within 14 days prior to the first ingestion of the investigational 
products to assess their eligibility to participate. For each child both parents will consent in 
writing (Appendix 1, Informed Consent Form) to the screening process before the start of the 
examination. The consent form will also include the study information leaflet.
The examinations and investigations will include:

Medical history, including history of past use of medications, demographics (date of birth, 
sex, race) and ASD diagnosis.

Physical examination including assessment of general appearance, cardiovascular, lungs, 
mouth, throat and abdomen and measurement of height and body weight. The examination 
will be made in accordance with the normal clinical routines at the trial center.

Vital signs: systolic and diastolic blood pressure in supine (after 5 minutes rest) and standing 
positions (1minute), pulse, respiratory rate and oral temperature.

Laboratory tests: HLA, Haematology, (see section 4.4.3.2); baseline plasma levels of IP, Els, 
LFts (these can be drawn also at V1 if considered by the study PI) 

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Based on the results of the testing with the M-CHAT-R, Q-CHAT and CAST questionnaires 
and the evaluation from child psychologist, a recommendation for treatment will be made for
either UCB administration or the noninvasive treatment for 6 weeks
Visit 1: The first treatment is administered

The pre-dose safety assessments and eligibility checks will be recorded on the relevant pages 
in the individual CRFs.

The parents will also be given the 3 questionnaires  and instructed on how to complete it.

The parents will be issued with prescriptions for trial medication, explained how to take the 
medication and given next appointment date; or

The UCB infusion is administered
Visit 2:  Second Treatment is administered
For these visits the examinations and investigations will include:

Physical examination (as for visit 1)

Vital signs: systolic and diastolic blood pressure in supine, pulse, respiratory rate and oral 
temperature.

Laboratory tests
HLA testing by genetic sequencing

Haematology: CBC - RBC, HB. MCHC, MCH, MCV, RDW, Hct, Hb, WBC 
Differential count –lymphocytes monocytes, neutrophils, eosinophils, basophils, 

flowcytometry - CD34+, TNC, viability %

Microbiology – presence and sensitivity of pathogens

LFTs Serum - Bilirubin (total and conjugated) AST, ALT, GGT, LDH, glucose

IP – serum protein electrophoresis, ESR, CRP, fibrinogen

Els - sodium, potassium, chloride, calcium, magnesium.
Note: For any patient in which any of the above tests indicated levels outside the normal range, a
follow-up visit (visit A-C) will be scheduled for an appropriate time (1 or 3 months post visit ). 
All or the above tests will be repeated at this visit.
4.3
Specific detail on measurements
4.4.1
Adverse events
An adverse event is the development of an undesirable medical condition  - e.g. symptoms or 
abnormal results of an investigation - or the deterioration of a pre-existing medical condition (not
relevant in this study).  AE’s will be collected by means of a standard question:  “Have you had 
any health problems since the previous visit?”  AE’s will be recorded at every visit. 
Spontaneously reported AE’s and/or observed AE’s and the subject’s response to this question 
will be recorded on the AE form with information about seriousness, action taken, date of onset 
and recovery, maximum intensity and outcome.  The subjects will be asked to assess the 
intensity of the reported Adverse Event according to the following scale:

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Mild 
    = awareness of sign or symptom, but easily tolerated
Moderate   = discomfort sufficient to cause interference with normal activities
Severe    =  incapacitating, with inability to perform normal activities.
A Serious Adverse Event is an adverse event occurring during any phase of the study and at any 
dose of the investigational product or placebo, which fulfils one or more of the following criteria:

Results in death;

Is immediately life-threatening;

Requires in-subject hospitalization; 

Results in persistent or significant disability or incapacity.
The causality of Serious Adverse Events (i.e. the relationship to study treatment) will be assessed
by the investigators, who in completing the relevant Case Report Form must answer ‘yes’ or ‘no’
to the question “Do you consider that there is a reasonable possibility that the event may have 
been caused by the study medication?”   The following factors should be considered when 
deciding if there is a “reasonable possibility” that an Adverse Event may have been caused by 
the investigational product.

Time course of events and exposure to suspect drug – did the AE occur in a reasonable 
temporal relationship to the administration of suspect drug?

Dechallenge experience – did the AE resolve or improve on stopping or reducing the dose of 
the suspect drug?

Rechallenge experience - did the AE reoccur if the suspected drug was reintroduced after 
having stopped?

Laboratory tests – has a specific laboratory investigation confirm the relationship?

No alternative cause -  the AE cannot be reasonably explained by another aetiology such as 
an underlying disease (not previously present), other drugs or environmental factors.
There would not be a “reasonable possibility” of causality if none of the above criteria apply or 
where there is evidence of exposure and a reasonable time course, but any dechallenge is 
negative or there is another more likely cause of the AE.
In this study the Adverse Events will be noted from the interview at the time of visit and from 
the daily input in the diary.  
4.4.2
12- Lead ECG
Standard 12-lead ECGs will be performed at the indication of the anesthesiology doctor if 
necessary.
The Bioclinica Laboratory will perform all the clinical laboratory investigations. 
The samples will be processed and shipped as per instructions of the pathology laboratory.
For all laboratory investigations the investigator will receive actual printouts of the data from the
Laboratory. A signed original printout will be attached to the CRF and a copy thereof will be 
filed in the subject file.  The reference ranges will be provided in the clinical trial manual 
(investigator’s file) together with the descriptions of the laboratory methods used. The laboratory
will provide normal ranges on each report to ease the investigator’s assessment.

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As part of the overall safety monitoring plan, the safety monitor (Dr Bogdan Ivanescu) will also 
assess the data, particularly those falling outside the normal range. 
The blood samples will always be taken before the first trial medication dose for that day is 
taken. The blood samples will be collected in tubes labeled with the following information: 
patient name; test type (HLA, biochemistry, hematology, flowcytometry); blood sample number 
(visit 2, 3,4 or 5) and sampling time.
Blood samples will be handled according to the protocols for testing of the respective 
laboratories
4.4
 Measurements recorded daily in a diary
A diary will be filled in at home during the entire study in the morning after taking the morning 
dose and at night after the evening before going to bed.  The following variables will be 
recorded:
i)
Intake of study medication (if, time and if with food) 
ii)
Occurrence of any adverse effects.
iii)
Intake of any incidental medication 
The subjects will record the time (morning and evening) that the study medication has been 
taken.  The recordings are a measurement of compliance.
5
STATISTICAL METHODS
5.1
Determination of sample size
Based on literature we expect that a sample size of minimum 25 participants will be needed in 
order to observe an alpha value of 0.05 (95% probability) with a p value of 0.05 and a power of 
80%. ANOVA testing will be performed on the respective groups of patients
5.2 Statistical analysis
A single statistical analysis will be performed at the end of the study. An intention to treat (ITT) 
approach will followed, i.e. statistical analysis of safety will be based on data from all patients 
and from whom meaningful data were collected. Data will be displayed graphically for visual 
inspection. Descriptive statistics will be presented as means, SEM and ninety percent confidence 
levels of the means.
Baseline characteristics. The subject disposition will be summarized. Protocol violations will 
be listed per subject, describing the nature of the violation. Subjects failing to complete the study
(as well as the times and reasons for discontinuation) will be displayed. The demographic, 
background and baseline data will be presented descriptively.

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Analysis of Safety.  Adverse Events, as reported throughout the course of the trial will be listed 
individually, per treatment group. Pre-, study and post-study findings of physical examination, 
vital sign variables, laboratory variables (haematology, clinical chemistry and urinalysis), and 
12-lead ECG if needed will be listed individually and summarized; values outside the normal 
range will be listed. 
.
5.3.
Changes to the Clinical Study Protocol
The Angiomedica Hospital (as the clinical site for the study) and the Local Ethics Committees 
will be notified of any amendments to the Clinical Study Protocol. Such changes will also be 
registered at the www.clinicaltrials.gov site
6. ETHICS
6.1
Ethics review
The final study protocol, including the final version of the Informed Consent Forms, must be 
approved in writing by The Ethics Committee before enrolment of any subject into the study.  
The Principal Investigator (Clinical Trial Manager) is responsible for informing the IEC of any 
serious adverse events (SAE) and amendment to the protocol as per regulatory requirement.
 6.2.
Ethical conduct of the study
The study will be performed in accordance with the ethical principles in the Declaration of 
Helsinki of the World Health Organisation (see Appendix 10.1), and that are consistent with 
Good Clinical Practice and applicable regulatory requirements.
6.3.
Subject information and consent
The Investigator will ensure that parents are given full and adequate oral and written information
about the nature, purpose, possible risk and benefit of the study.  Subjects must also be notified 
that they are free to discontinue from the study at any time.  The subject should be given the 
opportunity to ask questions and allowed time to consider the information provided.  The 
subject’s signed and dated informed consent must be obtained before conducting any study 
specific procedure. The investigator must store the original, signed Subject Informed Consent 
Form and a copy must be given to the subject.  Samples of the English version of the Subject 
Information and Consent Forms are enclosed (Appendix 1).  
6.4.
Subject data protection
The Subject Information and Consent Form will explain that study data will be stored in a 
computer database, maintaining confidentiality.  Subjects in this database will be identified by 
initials or enrolment code / subject number only.  Authorized representative of a regulatory 

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authority (e.g. ANM) may require direct access to parts of the trial site records relevant to the 
study, including subjects’ medical history for data verification purposes.
The Investigator must keep a Subject Identification List of all subjects that have signed the 
informed consent, including subject number, full name and last known address. 
7.
DATA QUALITY ASSURANCE
Data from the study will be collected in CRFs. Data editing will be performed at the trial center, 
comparing source and CRF entries. Data will be entered in a blind mode. 
During the study an independent monitor will visit the investigational site to confirm that the 
facilities remain acceptable, that the investigational team is adhering to the protocol and that data
are being accurately recorded in the CRFs.  Source data verification (a comparison of the data in 
the CRF with the subjects laboratory test results and other source documents) will also be 
performed.
Authorized representatives of the regulatory authority (e.g. ANM) may visit the center to 
perform inspections, including source data verification.
Clean File for the final database will be declared when all data have been entered and a quality 
check on a sample of the data has been performed.  The database will be locked after Clean File 
has been declared and data extracted for statistical analysis. Treatment code will not be broken 
until clean file. 
The medical, nursing and other staff involved in the study will receive proper 
education/information on how to conduct the study according to the protocol.
8.
STUDY TIME TABLE AND TERMINATION
First participant in 
Jan 2019
Last participant out 
March 2022
Clean File
March 2022
Study Report
April 2022
9.
REFERENCES
Braunschweig D, Duncanson P, Boyce R et al. Behavioral correlates of 
maternal antibody status among children with autism. J Autism Dev Disord 
2012;42:1435–1445
Courchesne E, Mouton PR, Calhoun ME, Semendeferi K, Ahrens-Barbeau C, 
Hallet MJ, Barnes CC, Pierce K. Neuron number and size in prefrontal cortex of
children with autism. JAMA. 2011 Nov 9; 306(18):2001-10
Croen LA, Braunschweig D, Haapanen L, et al. Maternal mid-pregnancy 

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autoantibodies to fetal brain protein: the early markers for autism study. Biol 
Psychiatry. 2008;64(7):583–588. doi:10.1016/j.biopsych.2008.05.006 
Dawson G, Sun JM, Davlantis KS, et al. Autologous Cord Blood Infusions Are 
Safe and Feasible   in Young Children   with Autism Spectrum Disorder: Results
of a Single-Center Phase I Open-Label Trial. Stem Cells Transl Med. 
2017;6(5):1332–1339. doi:10.1002/sctm.16-0474 
Geschwind DH, Levitt P. Autism spectrum disorders: developmental 
disconnection syndromes. Curr Opin Neurobiol. 2007 Feb; 17(1):103-11
Kornberg MD, Bhargava P, Kim PM, et al. Dimethyl fumarate targets GAPDH 
and aerobic glycolysis to modulate immunity. Science. 2018;360(6387):449–453.
doi:10.1126/science.aan4665
Nordahl CW, Lange N, Li DD, Barnett LA, Lee A, Buonocore MH, Simon TJ, 
Rogers S, Ozonoff S, Amaral DG. Brain enlargement is associated with 
regression in preschool-age boys with autism spectrum disorders. Proc Natl 
Acad Sci U S A. 2011 Dec 13; 108(50):20195-200.
Nordahl CW, Scholz R, Yang X, Buonocore MH, Simon T, Rogers S, Amaral DG.
Increased rate of amygdala growth in children aged 2 to 4 years with autism 
spectrum disorders: a longitudinal study. Arch Gen Psychiatry. 2012 Jan; 
69(1):53-61.
Pardo CA, Vargas  DL, Zimmerman AW. Immunity, neuroglia and 
neuroinflammation in autism.  Int Rev Psychiatry. 2005 Dec; 17(6):485-95.
Parker KJ, Oztan O, Libove RA, et al. Intranasal oxytocin treatment for social 
deficits and biomarkers of response in children with autism. Proc Natl Acad Sci
U S A. 2017;114(30):8119–8124. doi:10.1073/pnas.1705521114 
Sacco R, Militerni R, Frolli A, Bravaccio C, Gritti A, Elia M, Curatolo P, Manzi 
B, Trillo S, Lenti C, Saccani M, Schneider C, Melmed R, Reichelt KL, Pascucci T,
Puglisi-Allegra S, Persico AM. Clinical, morphological, and biochemical 
correlates of head circumference in autism. Biol Psychiatry. 2007 Nov 1; 
62(9):1038-47
Xydakis MS, Mulligan LP, Smith AB, Olsen CH, et al. Autism: Reduced 
Connectivity between Cortical Areas Involved with Face Expression, Theory of 
Mind, and the Sense of Self. Brain. Published online March 20 2015; 
doi:10.1093/brain/awv051 
Young AM, Chakrabarti B, Roberts D, Lai MC, Suckling J, Baron-Cohen S. 
From molecules to neural morphology: understanding neuroinflammation in 
autism spectrum condition. Mol Autism. 2016;7:9. Published 2016 Jan 20. 
doi:10.1186/s13229-016-0068-x 
 

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APPENDIX  A
DECLARATION OF HELSINKI
WORLD MEDICAL ASSOCIATION DECLARATION OF HELSINKI
Ethical Principles for Medical Research Involving Human Subjects 
Adopted by the 18th WMA General Assembly, Helsinki, Finland, June 1964, and amended by the
29th WMA General Assembly, Tokyo, Japan, October 1975
35th WMA General Assembly, Venice, Italy, October 1983
41st WMA General Assembly, Hong Kong, September 1989
48th WMA General Assembly, Somerset West, Republic of South Africa, October 1996
and the 52nd WMA General Assembly, Edinburgh, Scotland, October 2000 
Note of Clarification on Paragraph 29 added by the WMA General Assembly, Washington 2002
INTRODUCTION 
1.
The World Medical Association has developed the Declaration of Helsinki as a statement of 
ethical principles to provide guidance to physicians and other participants in medical research 
involving human subjects. Medical research involving human subjects includes research on 
identifiable human material or identifiable data.
2.
It is the duty of the physician to promote and safeguard the health of the people. The physician's 
knowledge and conscience are dedicated to the fulfillment of this duty.
3.
The Declaration of Geneva of the World Medical Association binds the physician with the words, 
"The health of my patient will be my first consideration," and the International Code of Medical 
Ethics declares that, "A physician shall act only in the patient's interest when providing medical 
care which might have the effect of weakening the physical and mental condition of the patient." 
4.
Medical progress is based on research, which ultimately must rest in part on experimentation 
involving human subjects.
5.
In medical research on human subjects, considerations related to the well-being of the human 
subject should take precedence over the interests of science and society.
6.
The primary purpose of medical research involving human subjects is to improve prophylactic, 
diagnostic and therapeutic procedures and the understanding of the aetiology and pathogenesis of 
disease. Even the best-proven prophylactic, diagnostic, and therapeutic methods must 
continuously be challenged through research for their effectiveness, efficiency, accessibility and 
quality. 
7.
In current medical practice and in medical research, most prophylactic, diagnostic and therapeutic 
procedures involve risks and burdens. 
8.
Medical research is subject to ethical standards that promote respect for all human beings and 
protect their health and rights. Some research populations are vulnerable and need special 
protection. The particular needs of the economically and medically disadvantaged must be 
recognized. Special attention is also required for those who cannot give or refuse consent for 
themselves, for those who may be subject to giving consent under duress, for those who will not 
benefit personally from the research and for those for whom the research is combined with care. 
9.
Research Investigators should be aware of the ethical, legal and regulatory requirements for 
research on human subjects in their own countries as well as applicable international requirements.

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No national ethical, legal or regulatory requirement should be allowed to reduce or eliminate any 
of the protections for human subjects set forth in this Declaration.
B.
BASIC PRINCIPLES FOR ALL MEDICAL RESEARCH 
10. It is the duty of the physician in medical research to protect the life, health, privacy, and dignity of 
the human subject. 
11. Medical research involving human subjects must conform to generally accepted scientific 
principles, be based on a thorough knowledge of the scientific literature, other relevant sources of 
information, and on adequate laboratory and, where appropriate, animal experimentation.
12. Appropriate caution must be exercised in the conduct of research, which may affect the 
environment, and the welfare of animals used for research must be respected.
13. The design and performance of each experimental procedure involving human subjects should be 
clearly formulated in an experimental protocol. This protocol should be submitted for 
consideration, comment, guidance, and where appropriate, approval to a specially appointed 
ethical review committee, which must be independent of the investigator, the sponsor or any other 
kind of undue influence. This independent committee should be in conformity with the laws and 
regulations of the country in which the research experiment is performed. The committee has the 
right to monitor ongoing trials. The researcher has the obligation to provide monitoring 
information to the committee, especially any serious adverse events. The researcher should also 
submit to the committee, for review, information regarding funding, sponsors, institutional 
affiliations, other potential conflicts of interest and incentives for subjects. 
14. The research protocol should always contain a statement of the ethical considerations involved and
should indicate that there is compliance with the principles enunciated in this Declaration. 
15. Medical research involving human subjects should be conducted only by scientifically qualified 
persons and under the supervision of a clinically competent medical person. The responsibility for 
the human subject must always rest with a medically qualified person and never rest on the subject
of the research, even though the subject has given consent. 
16. Every medical research project involving human subjects should be preceded by careful 
assessment of predictable risks and burdens in comparison with foreseeable benefits to the subject 
or to others. This does not preclude the participation of healthy volunteers in medical research. 
The design of all studies should be publicly available.
17. Physicians should abstain from engaging in research projects involving human subjects unless 
they are confident that the risks involved have been adequately assessed and can be satisfactorily 
managed. Physicians should cease any investigation if the risks are found to outweigh the potential
benefits or if there is conclusive proof of positive and beneficial results. 
18. Medical research involving human subjects should only be conducted if the importance of the 
objective outweighs the inherent risks and burdens to the subject. This is especially important 
when the human subjects are healthy volunteers. 
19. Medical research is only justified if there is a reasonable likelihood that the populations in which 
the research is carried out stand to benefit from the results of the research. 
20. The subjects must be volunteers and informed participants in the research project.
21. The right of research subjects to safeguard their integrity must always be respected. Every 
precaution should be taken to respect the privacy of the subject, the confidentiality of the patient's

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information and to minimize the impact of the study on the subject's physical and mental integrity 
and on the personality of the subject.
22. In any research on human beings, each potential subject must be adequately informed of the aims, 
methods, sources of funding, any possible conflicts of interest, institutional affiliations of the 
researcher, the anticipated benefits and potential risks of the study and the discomfort it may 
entail. The subject should be informed of the right to abstain from participation in the study or to 
withdraw consent to participate at any time without reprisal. After ensuring that the subject has 
understood the information, the physician should then obtain the subject's freely given informed 
consent, preferably in writing. If the consent cannot be obtained in writing, the non-written 
consent must be formally documented and witnessed. 
23. When obtaining informed consent for the research project the physician should be particularly 
cautious if the subject is in a dependent relationship with the physician or may consent under 
duress. In that case the informed consent should be obtained by a well-informed physician who is 
not engaged in the investigation and who is completely independent of this relationship. 
24. For a research subject who is legally incompetent, physically or mentally incapable of giving 
consent or is a legally incompetent minor, the investigator must obtain informed consent from the 
legally authorized representative in accordance with applicable law. These groups should not be 
included in research unless the research is necessary to promote the health of the population 
represented and this research cannot instead be performed on legally competent persons. 
25. When a subject deemed legally incompetent, such as a minor child, is able to give assent to 
decisions about participation in research, the investigator must obtain that assent in addition to the 
consent of the legally authorized representative. 
26. Research on individuals from whom it is not possible to obtain consent, including proxy or 
advance consent, should be done only if the physical/mental condition that prevents obtaining 
informed consent is a necessary characteristic of the research population. The specific reasons for 
involving research subjects with a condition that renders them unable to give informed consent 
should be stated in the experimental protocol for consideration and approval of the review 
committee. The protocol should state that consent to remain in the research should be obtained as 
soon as possible from the individual or a legally authorized surrogate.
27. Both authors and publishers have ethical obligations. In publication of the results of research, the 
investigators are obliged to preserve the accuracy of the results. Negative as well as positive 
results should be published or otherwise publicly available. Sources of funding, institutional 
affiliations and any possible conflicts of interest should be declared in the publication. Reports of 
experimentation not in accordance with the principles laid down in this Declaration should not be 
accepted for publication. 
C.
ADDITIONAL PRINCIPLES FOR MEDICAL RESEARCH COMBINED WITH MEDICAL CARE
28. The physician may combine medical research with medical care, only to the extent that the 
research is justified by its potential prophylactic, diagnostic or therapeutic value. When medical 
research is combined with medical care, additional standards apply to protect the patients who are 
research subjects.
29. The benefits, risks, burdens and effectiveness of a new method should be tested against those of 
the best current prophylactic, diagnostic, and therapeutic methods. This does not exclude the use 
of placebo, or no treatment, in studies where no proven prophylactic, diagnostic or therapeutic 
method exists. See footnote 

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30. At the conclusion of the study, every patient entered into the study should be assured of access to 
the best-proven prophylactic, diagnostic and therapeutic methods identified by the study.
31. The physician should fully inform the patient which aspects of the care are related to the research. 
The refusal of a patient to participate in a study must never interfere with the patient-physician 
relationship.
32. In the treatment of a patient, where proven prophylactic, diagnostic and therapeutic methods do 
not exist or have been ineffective, the physician, with informed consent from the patient, must be 
free to use unproven or new prophylactic, diagnostic and therapeutic measures, if in the 
physician's judgement it offers hope of saving life, re-establishing health or alleviating suffering. 
Where possible, these measures should be made the object of research, designed to evaluate their 
safety and efficacy. In all cases, new information should be recorded and, where appropriate, 
published. The other relevant guidelines of this Declaration should be followed. 
FOOTNOTE:
NOTE OF CLARIFICATION ON PARAGRAPH 29 of the WMA DECLARATION OF HELSINKI 
The WMA hereby reaffirms its position that extreme care must be taken in making use of a placebo-controlled trial 
and that in general this methodology should only be used in the absence of existing proven therapy. However, a 
placebo-controlled trial may be ethically acceptable, even if proven therapy is available, under the following 
circumstances:
- Where for compelling and scientifically sound methodological reasons its use is necessary to determine the 
efficacy or safety of a prophylactic, diagnostic or therapeutic method; or 
- Where a prophylactic, diagnostic or therapeutic method is being investigated for a minor condition and the patients
who receive placebo will not be subject to any additional risk of serious or irreversible harm. 
All other provisions of the Declaration of Helsinki must be adhered to, especially the need for appropriate ethical 
and scientific review.
The Declaration of Helsinki (Document 17.C) is an official policy document of the World Medical Association, the 
global representative body for physicians. It was first adopted in 1964 (Helsinki, Finland) and revised in 1975 
(Tokyo, Japan), 1983 (Venice, Italy), 1989 (Hong Kong), 1996 (Somerset-West, South Africa) and 2000 
(Edinburgh, Scotland). Note of clarification on Paragraph 29 added by the WMA General Assembly, Washington 
2002.
 

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APPENDIX 1
PATIENT INFORMATION LEAFLET AND INFORMED CONSENT
PATIENT INFORMATION SHEET AND 
PATIENT INFORMATION SHEET AND 
INFORMED CONSENT FORM
INFORMED CONSENT FORM
Title of the study:  
A pilot crossover study to investigate the safety and efficacy  of
administering intravenous autologous cord blood versus personalized
noninvasive treatments to children with developmental disorders of
the autistic spectrum.
Protocol number: 
B-F011
Sponsor: 
The Bio-Forum Foundation
Name of Principal Investigator: 
Dr Felician Stancioiu
Name of Study Coordinator:
Dr Bogdan Ivanescu
Address of site/institution where study will be conducted:  
Spitalul Angiomedica, Str. Cosminului Nr, 13, Bucuresti
INTRODUCTION 
You are being invited to take part in a clinical research study sponsored by The  Bio-Forum
Foundation. Bio-Forum Foundation is engaged in joint research with Angiomedica Hospital on stem
cell treatments. Before deciding to take part in the study, it is important for you to understand why
the research is done and what will happen to participants in the study. Please take the time to read
this information carefully and ask the investigator when anything is not clear, or if you would like
more information.
What is the purpose of this study?
The purpose of this study is to determine the comparative safety and efficacy of the umbilical cord
blood versus noninvasive treatments. 
All participants will be administered both treatments (umbilical cord blood and the noninvasive
treatment).
Do I have to participate in the study?
Participation in this study is entirely voluntary. It is up to you to decide whether to take part or not.
If you do not want to take part in this study or if you wish to withdraw from the study at any time you
may do so without giving a reason and you will not lose any benefits to which you would otherwise
be entitled. If you withdraw from the study the data collected up to the point of withdrawal will be
analyzed for the purpose of the study. If you decide to participate, you will be given this information
sheet to keep and be asked to provide your signature indicating your consent.
Patient identification number: . . . .  / . . . . .                     Patient initials: 

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Who is eligible to participate in the study?
About 25-40 children age 3-7 years will take part in this study.  In order to participate in this trial, the
child needs to be diagnosed with ASD, be in the appropriate weight range, not on any regular
medicine,  and  meet  the  inclusion  criteria  of  the  study,  which  will  be  told  by  the  principal
investigator. 
 
Safety of medications is discussed for each of the recommended treatments at time of enrollment in
the study.
What will happen to me if I participate?
If you decide to take part you must sign the consent form and will then first be examined to see if you
meet all the requirements for participation in the study.  These examinations and investigations (at 
visit 1) will include the taking of a medical history history, including history of past use of 
medications; a physical examination; the withdrawal of 40 ml (approximately 8 teaspoons) of blood 
for laboratory tests; and other tests as deemed appropriate for each patient.
If you qualify for inclusion you will thereafter, within 21 days of the first examination, at Visit 1
will either be prescribed study medication that you must take for 6 weeks or prepare for UCB
infusion which will take place at the Angiomedica Hospital.
You will have to take the study medication for a total of 6 weeks and return for visits to the trial
center for a further 3 visits, at 16, 24 and 52 weeks. 
If UCB infusion is given first, then at 8 weeks after UCB infusion, the child will start taking the
non-invasive treatment recommended after the evaluations.
This means that the study will last around 12 months.
What do I have to do?
As a subject in this study, you are responsible for:

keeping all clinic appointments; 

taking the study medication as explained by the study doctor;

completing and returning the questionnaires that you will receive at each study visit;

telling your study doctor before taking any other medication;

telling your study doctor as soon as possible if there is a change in your health;
 
If you experience any injuries and need to see a doctor, you must also tell that doctor that you are 
participating in this research study.
What risks or discomforts might occur if I participate?
As with any medication and treatment, there may be side-effects associated with the administered 
therapies, the most common of which is cited in literature – episodes of agitation seen after UCB 
administration
Some procedures used in this study may cause some discomforts (e.g. the withdrawal of blood may
cause  slight  bruising  or  possible  fainting,  there  may  be  discomfort  during  infusion  or  after
administration of sedation, etc), but these will be no more than that experienced in a typical medical
examination. 
What are the possible benefits of participating?
You may or may not receive any direct benefit from the two treatments, the only guarantee is that the
information we get from this study will extend our existing knowledge and will help to decide if this
medication is safe to use in other people such as you.

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Will there be any cost to me if I participate?
For participating in this research study, you will need to cover the costs for the materials,
medications, consumables, hospital stay, UCB preparation and infusion. testing, clinic visits; the
total cost is estimated at between 4.500 and 5000 euros, which will be paid to Fundatia Bio-
Forum in two equal installments, half (2000-2500) at enrollment in the clinical study and the
remainder half at UCB treatment completion (8 weeks after first visit).
What if something goes wrong or if I have problems while I am in the study?
In case of study related questions please contact:
Doctor:
Dr Felician Stancioiu    or     Dr. Bogdan Ivanescu
Contact Numbers:
0727500402                   at           0722504540
If you are harmed by your participation in the study, you will be compensated according to the
guidelines of the pharmaceutical industry relating to clinical trials (Association for British
Pharmaceutical Industry ABPI guidelines). If you are harmed due to someone’s negligence then
you may have grounds for legal action. You are not waiving your legal rights by signing this
form.
Can I withdraw or be withdrawn from the study? 
Taking part in the study is voluntary. If you decide to take part in the study, you are free to
withdraw from the study at any time.  If you decide to withdraw from the study, you should
inform your study doctor immediately.  Your study doctor will not be upset and you will not be
penalized in any way, and your future care will not be affected.  Should you withdraw from the
study, the study data collected before your withdrawal may still be processed along with other
data collected as part of the study.
In addition, circumstances may arise that will lead to the ending of your participation in this
study. Such circumstances could include:

not taking your medication or not regularly keeping to the study appointments, or if you do 
not follow the doctor's instructions;

taking a medication that interferes with the treatments in this study (eg steroids, immune 
supressant, benzodiazepine, other psychotropic medication);

having blood tests which are outside of normal

if the Ethical Committee of Fundatia Bio-Forum stops or suspends the study.
Will my participation in this study be kept confidential?
The records that identify participants will be kept confidential and, to the extent permitted by the 
applicable laws and regulations, will not be made publicly available. If you consent to take part 
in this study, any of the medical records and data recorded in this study may be directly inspected
by representatives of the study Sponsor (Bio-Forum Foundation), Stem Cell Banks and 
Angiomedica Hospital. By signing this written informed consent form you are giving permission 
for this to be done. 
The information collected during the study will be stored in a computer but your name will not
be stored. Only your study doctor will know that the information is related to you.  

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The results of the study may be published in the medical literature and/or presented at a scientific
conference or symposium, but your identity will not be revealed. The information disclosed will
be collective summarized data.
You may ask to see your medical information as prescribed by law.
Who is organising and funding the study?
This  research  study  is  being  organized  and  funded  by  Bio-Forum  Foundation  in  close
collaboration with Angiomedica Hospital and Stem Cell Banks. Dr Felician Stancioiu is the
clinical investigator responsible for the conduct of the trial and the contact person for Bio-Forum
Foundation and Dr Bogdan Ivanescu, the Study Coordinator. 
Who has reviewed the study? 
The study is in accordance with the guidelines of the International Conference on Harmonization
(ICH) for Good Clinical Practice (GCP) and with the Declaration of Helsinki (version 2000).This
study has been approved by the Research Ethics Committee. The study will also be reviewed by
the Institutional Review Board of the Angiomedica Hospital (partner in the study with Bio-
Forum Foundation). These are groups of scientific and non-scientific people. They review and
approve or disapprove research involving human subjects.  
 
You should inform your general practitioner (your house doctor) that you are taking part in this
study.    
Contacts for further information.
If during the course of this study, you have questions about the nature of the research or your
rights, or you believe that you have sustained a research-related injury, you should contact one of
the following: 
Dr Felician Stancioiu (study doctor) at 0727500402  
 

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INFORMED CONSENT FORM
Clinical study title:   A pilot crossover study to investigate the safety and efficacy of
administering  intravenous  autologous  cord  blood  versus  personalized
noninvasive  treatments  to children  with  developmental  disorders  of  the
autistic spectrum
Protocol number: B-F001
By signing and dating this document,

I confirm that I have had time to carefully read and understand the patient information sheet 
provided for this study.

I confirm that I have had the opportunity to discuss the study and ask questions and I am satisfied
with the answers and explanations that I have been provided.

I give permission for my medical records to be reviewed by the Sponsor or designee, and/or 
representatives of the Medicines Control Council or Committee for Pharmaceutical Trials.

I understand that my participation is voluntary and that I am free to withdraw at any time without
giving any reason and without my medical care or legal rights being affected.

I confirm that I have received a signed and dated copy of the Patient Information Sheet and 
Informed Consent Forms.
Child’s Name, Mother: 
_________________  _______________    ______________
      
Name (capital letters)
Signature
     Date
Father:                  
 _________________   _______________   _________________
Name (capital letters)             Signature                          Date
PERSON OBTAINING CONSENT:
     
_________________  _______________    ________________
      
Name (capital letters)
Signature
     Date
CLINICAL INVESTIGATOR:
     
_________________  _______________    ________________
      
Name (capital letters)
Signature
     Date
Patient identification number:      . . . .  / . . . . .       Patient initials: 

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ANNEX 1 
COSTS
Infusion of the UCB collection 
           
4500 euro
includes 
- transport and storage of UCB in liquid nitrogen (Bio-Forum, Angiomedica)
- HLA testing of the child (National Institute of Hematology Lab)
- laboratory testing - CBC, LFTs, Els, IP (Bioclinica Lab)
- HLA testing of the UCB (National Institute of Hematology Lab)
- thawing, conditioning of UCB (Bio-Forum, Angiomedica)
- Trypan Blue and microbiological testing (Angiomedica and Bioclinica Lab)
- flowcytometry (TNC and viability) – Institutul de Biochimie al Academiei 
- preparation of UCB collection for infusion according to Vita34 protocols (Bio-
Forum, Angiomedica)
- hospital admission (24 hours), pre-medication, administration of UCB by 
anesthesiologist (Angiomedica)
- other testing (EKG, blood, etc) and medical care, as needed (Angiomedica)
Also included:
- interview/recruitment by study coordinator (Stem Cell Banks)
- evaluation by child therapist/psychologist (4 visits)
- M-CHAT/Q-CHAT/CAST questionnaires
- evaluation by study investigator and prescription of supplements for 6 weeks
- a second set of blood tests (CBC, LFT, El, IP)
Additional possible, optional costs
- MRI at 9-12 months after UCB infusion (Monza Hospital),                       400 euro
- evaluation by another child therapist (per visit)                                     50 euro
- Additional follow-up visits, 3rd set of blood tests 
               900 euro

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Annex 2
CAST Questionnaire
The Childhood Autism Spectrum Test (CAST)
Child’s Name: .................................. Age: ......................... Sex: Male / Female
Birth Order: ..................................... Twin or Single Birth: ..................................
Parent/Guardian: .....................................................................................................
Parent(s) occupation: ............................................................................................
Age parent(s) left full-time education: ....................................................................
Address: .................................................................................................................
 .................................................................................................................
.................................................................................................................
Tel.No: .................................. School: ........................................................
----------------------------------------------------------------------------------------------------
Please read the following questions carefully, and circle the appropriate answer. All responses are confidential.
1. Does s/he join in playing games with other children easily?  Yes No
2. Does s/he come up to you spontaneously for a chat? Yes No
3. Was s/he speaking by 2 years old? Yes No
4. Does s/he enjoy sports? Yes No
5. Is it important to him/her to fit in with the peer group? Yes No
6. Does s/he appear to notice unusual details that  others miss? Yes No
7. Does s/he tend to take things literally? Yes No
8. When s/he was 3 years old, did s/he spend a lot of time  pretending (e.g., play-acting being a superhero, or
 holding teddy’s tea parties)? Yes No
9. Does s/he like to do things over and over again,  in the same way all the time? Yes No
10. Does s/he find it easy to interact with other  children? Yes No
11. Can s/he keep a two-way conversation going? Yes No
12. Can s/he read appropriately for his/her age? Yes No
13. Does s/he mostly have the same interests as  his/her peers? Yes No
14. Does s/he have an interest which takes up so much  time that s/he does little else? Yes No
15. Does s/he have friends, rather than just acquaintances? Yes No
16. Does s/he often bring you things s/he is interested  in to show you? Yes No
17. Does s/he enjoy joking around? Yes No
18. Does s/he have difficulty understanding the rules  for polite behaviour? Yes No
19. Does s/he appear to have an unusual memory for  details? Yes No

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20. Is his/her voice unusual (e.g., overly adult, flat, or  very monotonous)? Yes No
21. Are people important to him/her? Yes No
22. Can s/he dress him/herself? Yes No
23. Is s/he good at turn-taking in conversation? Yes No
24. Does s/he play imaginatively with other  children, and engage in role-play? Yes No
25. Does s/he often do or say things that are tactless  or socially inappropriate? Yes No
26. Can s/he count to 50 without leaving out any  numbers? Yes No
27. Does s/he make normal eye-contact? Yes No
28. Does s/he have any unusual and repetitive  movements? Yes No
29. Is his/her social behaviour very one-sided and  always on his/her own terms? Yes No
30. Does s/he sometimes say “you” or “s/he” when  s/he means “I”? Yes No
31. Does s/he prefer imaginative activities such as  play-acting or story-telling, rather than numbers  or lists of facts?
Yes No
32. Does s/he sometimes lose the listener because of  not explaining what s/he is talking about? Yes No
33. Can s/he ride a bicycle (even if with stabilisers)? Yes No
34. Does s/he try to impose routines on him/herself,  or on others, in such a way that it causes problems? Yes No
35. Does s/he care how s/he is perceived by the rest of  the group? Yes No
36. Does s/he often turn conversations to his/her  favourite subject rather than following what the other
 person wants to talk about? Yes No
37. Does s/he have odd or unusual phrases? Yes No
SPECIAL NEEDS SECTION
Please complete as appropriate
38. Have teachers/health visitors ever expressed any  concerns about his/her development? Yes No
If Yes, please specify..................................................................................................
39. Has s/he ever been diagnosed with any of the following?:
Language delay Yes No
Hyperactivity/Attention Deficit Disorder (ADHD) Yes No
Hearing or visual difficulties Yes No
Autism Spectrum Condition, incl. Asperger’s Syndrome Yes No
A physical disability Yes No
Other (please specify) Yes No

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Annex 3
M-CHAT-R Questionnaire
www.m-chat.org
Childʼs name                                                                                   Date
Age                                             Relationship to child
M-CHAT-RTM (Modified Checklist for Autism in Toddlers Revised)
Please answer these questions about your child. Keep in mind how your child usually behaves. If you have seen your
child do the behavior a few times, but he or she does not usually do it, then please answer no. Please circle yes or no 
for every question. Thank you very much.
1. If you point at something across the room, does your child look at it? Yes No
(FOR EXAMPLE, if you point at a toy or an animal, does your child look at the toy or animal?)
2. Have you ever wondered if your child might be deaf? Yes No
3. Does your child play pretend or make-believe?  Yes  No
(FOR EXAMPLE, pretend to drink from an empty cup, pretend to talk on a phone, or pretend to feed a doll or 
stuffed animal?)
4. Does your child like climbing on things? Yes   No (FOR EXAMPLE, furniture, playground equipment, or stairs)
5. Does your child make unusual finger movements near his or her eyes? Yes No
(FOR EXAMPLE, does your child wiggle his or her fingers close to his or her eyes?)
6. Does your child point with one finger to ask for something or to get help? Yes No
(FOR EXAMPLE, pointing to a snack or toy that is out of reach)
7. Does your child point with one finger to show you something interesting? Yes No
(FOR EXAMPLE, pointing to an airplane in the sky or a big truck in the road)
8. Is your child interested in other children? (FOR EXAMPLE, does your child watch Yes No
other children, smile at them, or go to them?)
9. Does your child show you things by bringing them to you or holding them up for you to Yes No
see – not to get help, but just to share? (FOR EXAMPLE, showing you a flower, a stuffed
animal, or a toy truck)
10. Does your child respond when you call his or her name? (FOR EXAMPLE, does he or she
look up, talk or babble, or stop what he or she is doing when you call his or her name?)
Yes No
11. When you smile at your child, does he or she smile back at you? Yes No
12. Does your child get upset by everyday noises? (FOR EXAMPLE, does your
child scream or cry to noise such as a vacuum cleaner or loud music?)
Yes No
13. Does your child walk? Yes No
14. Does your child look you in the eye when you are talking to him or her, playing with him Yes No
or her, or dressing him or her?

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15. Does your child try to copy what you do? (FOR EXAMPLE, wave bye-bye, clap, or Yes No
make a funny noise when you do)
16. If you turn your head to look at something, does your child look around to see what you Yes No
are looking at?
17. Does your child try to get you to watch him or her? (FOR EXAMPLE, does your child Yes No
look at you for praise, or say “look” or “watch me”?)
18. Does your child understand when you tell him or her to do something? Yes No
(FOR EXAMPLE, if you don’t point, can your child understand “put the book
on the chair” or “bring me the blanket”?)
19. If something new happens, does your child look at your face to see how you feel about it? Yes No
(FOR EXAMPLE, if he or she hears a strange or funny noise, or sees a new toy, will
he or she look at your face?)
20. Does your child like movement activities? Yes No
(FOR EXAMPLE, being swung or bounced on your kne
Permissions for Use of the M-CHAT-R/FTM
The Modified Checklist for Autism in Toddlers, Revised with Follow-Up (M-CHAT-R/F; Robins, Fein, &
Barton, 2009) is a 2-stage parent-report screening tool to assess risk for Autism Spectrum Disorder
(ASD). The M-CHAT-R/F is available for free download for clinical, research, and educational purposes.
Download of the M-CHAT-R/F and related material is authorized from www.mchatscreen.com.
The M-CHAT-R/F is a copyrighted instrument, and use of the M-CHAT-R/F must follow these guidelines:
(1) Reprints/reproductions of the M-CHAT-R must include the copyright at the bottom ( 2009
Robins, Fein, & Barton). No modifications can be made to items, instructions, or item order
without permission from the authors.
(2) The M-CHAT-R must be used in its entirety. Evidence indicates that any subsets of items do not
demonstrate adequate psychometric properties.
(3) Parties interested in reproducing the M-CHAT-R/F in print (e.g., a book or journal article) or
electronically for use by others (e.g., as part of digital medical record or other software packages)
must contact Diana Robins to request permission (DianaLRobins@gmail.com).
(4) If you are part of a medical practice, and you want to incorporate the first stage M-CHAT-R
questions into your own practice’s electronic medical record (EMR), you are welcome to do so.
However, if you ever want to distribute your EMR page outside of your practice, please contact
Diana Robins to request a licensing agreement.

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Annex 4
Q-CHAT Questionnaire
Please answer the following questions about your child by ticking the appropriate circle.
Try to answer EVERY question if you can. 
1. Does your child look at you when you call his/her name?
○ always
○ usually
○ sometimes
○ rarely
○ never
2. How easy is it for you to get eye contact with your
child?
○ very easy
○ quite easy
○ quite difficult
○ very difficult
○ impossible
3. When your child is playing alone, does s/he line objects
up?
○ always
○ usually
○ sometimes
○ rarely
○ never
4. Can other people easily understand your child’s speech?
○ always
○ usually
○ sometimes
○ rarely
○ never
○ my child does not speak
5. Does your child point to indicate that s/he wants
something (e.g. a toy that is out of reach)?
○ many times a day
○ a few times a day
○ a few times a week
○ less than once a week
○ never
Please answer the following questions about your child by ticking the appropriate circle.
Try to answer EVERY question if you can.
6. Does your child point to share interest with you (e.g.
pointing at an interesting sight)?
○ many times a day
○ a few times a day
○ a few times a week
○ less than once a week
○ never 
7. How long can your child’s interest be maintained by a spinning object
(e.g. washing machine, electric fan, toy car wheels)?
○ several hours
○ half an hour
○ ten minutes
○ a couple of minutes

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○ less than a minute
8. How many words can your child say?
○ none—s/he has not started speaking yet
○ less than 10 words
○ 10–50 words
○ 51–100 words
○ over 100 words
9. Does your child pretend (e.g. care for dolls, talk on a toy
phone)?
○ many times a day
○ a few times a day
○ a few times a week
○ less than once a week
○ never
10. Does your child follow where you’re looking?
○ many times a day
○ a few times a day
○ a few times a week
○ less than once a week
○ never
11. How often does your child sniff or lick unusual
objects?
○ many times a day
○ a few times a day
○ a few times a week
○ less than once a week
○ never
12. Does your child place your hand on an object when s/he
wants you to use it (e.g. on a door handle when s/he wants you to
open the door, on a toy when s/he wants you to activate it)?
○ many times a day
○ a few times a day
○ a few times a week
○ less than once a week
○ never
13. Does your child walk on tiptoe?
○ always
○ usually
○ sometimes
○ rarely
○ never
14. How easy is it for your child to adapt when his/her
routine changes or when things are out of their usual
place?
○ very easy
○ quite easy
○ quite difficult
○ very difficult
○ impossible
15. If you or someone else in the family is visibly upset, does
your child show signs of wanting to comfort them
(e.g. stroking their hair, hugging them)?
○ always
○ usually
○ sometimes
○ rarely
○ never

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16. Does your child do the same thing over and
over again (e.g. running the tap, turning the light
switch on and off, opening and closing doors)?
○ many times a day
○ a few times a day
○ a few times a week
○ less than once a week
○ never
17. Would you describe your child’s first words
as:
○ very typical
○ quite typical
○ slightly unusual
○ very unusual
○ my child doesn’t speak
18. Does your child echo things s/he hears (e.g. things
that you say, lines from songs or movies, sounds)?
○ many times a day
○ a few times a day
○ a few times a week
○ less than once a week
○ never
19. Does your child use simple gestures (e.g. wave goodbye)?
○ many times a day
○ a few times a day
○ a few times a week
○ less than once a week
○ never
20. Does your child make unusual finger movements near his/her eyes?
○ many times a day
○ a few times a day
○ a few times a week
○ less than once a week
○ never
21. Does your child spontaneously look at your face to
check your reaction when faced with something
unfamiliar?
○ always
○ usually
○ sometimes
○ rarely
○ never
22. How long can your child’s interest be maintained by
just one or two objects?
○ most of the day
○ several hours
○ half an hour
○ ten minutes
○ a couple of minutes
23. Does your child twiddle objects
repetitively (e.g. pieces of string)?
○ many times a day
○ a few times a day
○ a few times a week
○ less than once a week
○ never
24. Does your child seem oversensitive to noise?

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○ always
○ usually
○ sometimes
○ rarely
○ never
25. Does your child stare at nothing with no apparent
purpose?
○ many times a day
○ a few times a day
○ a few times a week
○ less than once a week
○ never
Annex 5

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UCB thawing protocol (adapted from Duke/Rubinstein protocols and Vita 34)
Form FL.03.01.07, Version 1 Printed from StemCare QMS 2019-01-22 
 
CBU Thawing Procedures
Principles
Cryopreserved CBU is protected against freezing damage by the addition of a
cryoprotector (DMSO) and dextrane. Immediately after thawing the addition of a dextrane
and albumin solution restores the osmolarity of the blood cell suspension. The additional 
removal of the cryoprotector DMSO may be beneficial in transplant patients because when
given in high volumes (more than 30 ml or 1 ml/kg body weight) and/or concentrations 
above 10% DMSO can cause adverse effects in vivo after reinfusion especially in 
immunosupressed patients.
The most important principles in preparing the thawed CBU for infusion are:
1. least manipulation of the cells by using a minimum number of centrifugations steps while
recovering the greatest number of stem cells (both hematopoietic and mesenchymal); this 
ensures good viability, number and type of cells
2. lack of contamination/ensuring sterile conditions for the cell suspension which will be 
administered to the patient
The two principles will be observed simultaneously and with equal importance.
Specimen
Cryopreserved umbilical cord blood that has been designated for administration to a
particular recipient.
Equipment
Laminar Flow Hood
Centrifuge
Plasma Expressor
Digital Balance
Tube Sealer compatible for PVC tubing
Automated Cell Counter or Microscope and Chamber for counting of cells
Waterbath (4 litres or more at 38°C)
Reagents
Albumin (5% human USP)
10% Dextran 40 (Gentran 40) and 0.9% NaCl, USP
Methyl violet acetic acid, Türk’s solution, Leukoplate (or equivalent) for staining of cells
Supplies
Cell Wash/Infusion Bag 
If there is ANY error or ambiguity, advise StemCare and the Transplant team. Do not proceed 
until the problem is resolved. If your LN2 storage tanks have no space to store the unit in its 
canister, add LN2 to the Dry Shipper container to maintain the unit frozen until a complete 
satisfactory determination is made.
1. Transfer the CBU unit from the Dry Shipper container from liquid nitrogen to the vapor phase 
of a liquid nitrogen container.
2. Visualize the identity of the CBU unit (directly on the carbdoard or aluminum box o

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3. Check the information sheet submitted with the unit against the patient records to verify ID 
number
4. Check the unit number by examining the labels on the unit; both the bar-coded and human 
readable printed numbers should conform to the information in the form provided with the CBU 
unit and the patient records.
5. Leave the CBU in the vapor phase for 5-10 minutes before proceeding.
6. Prepare the thawing solutions as described in point 16 and 21.
7. Open the cardboard box. Work carefully to avoid damaging the frozen plastic bag.
Remember that plastic at this temperature is very brittle and breaks easily.
8. Remove the CBU unit from the canister.
Note: Do not allow the unit to bend or it will crack!
9. Place the unit inside a zipper-locked plastic bag, let the air out and close the bag. Put the
unit in a water bath at 38°C. Take care not to break the plastic tube segments, if still
attached.
10. To accelerate thawing, carefully move the unit in the water and knead its contents gently but 
continuously.
Note: Inspect for leaks. Plastic bags for frozen blood may have pin-holes, other defects or
develop leaks. It is most important to prevent serious blood losses or bacterial contamination 
caused by cracks or leaks. If blood leaks out into the zipper-locked bag, find the site of the break 
and position the unit so as to prevent further escape of blood. While maintaining the position, 
thaw the unit.
11. As soon as the blood has thawed, transfer the bag from the water bath into a laminar flow 
hood.
12. Remove the unit from the zipper-locked bag. Using an alcohol or iodine swab disinfect
the covers of both ports of the freezing bag.
13. With scissors, cut off both the hermetically sealed covers to the spike ports on the
freezing bag. Disinfect the scissors before using.
14. Using an alcohol or iodine swab, disinfect the cut surfaces of the spike port area of the
freezing bag.
15. Close all clamps on the Transplant/washing set. Insert the spikes of the transplant set in
the ports of the freezing bag.
Note: Label the transplant bag with the unique CBU unit number and the name of the
recipient, or according to local standard practice!
16. Fit an 18 gauge needle to a 50 ml syringe. Draw 12.5 ml of 10% Dextran 40 and 12.5 ml
of 5% human albumin into the syringe. This is the thawing solution and can be prepared
before thawing the CBU.
17. Attach the syringe with thawing solution to the female luer lock on the Transplant set.
Open pinch clamp 1, 2 and 3 (PC-1, PC-2 and PC-3), and then slowly introduce the 25 ml of 
dextran/albumin solution to the 25 ml CBU in the freezing bag. Mix the fluids during transfer. 
Close PC-3.
18. Allow 5 minutes for equilibration.
19. Open PC-4. Transfer the diluted CBU from the freezing bag to the transplant bag.
20. Pass the diluted CBU back and forth between the transplant bag and the freezing bag in order
to more completely wash all cells out of the freezing bag into the transplant bag. Close PC-1 and 
PC-2.
21. Fit a 18 gauge needle to a 60 ml syringe. Draw 30 ml of 10% Dextran 40 and 30 ml of 5% 
human albumin into the syringe. Prepare 4 x 60 ml syringes in total as described. The

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syringes can be prepared before thawing the CBU.
22. Attach a 60 ml syringe with dextran/albumin solution to the luer lock. Open PC-3.
Transfer the 60 ml solution to the diluted CBU in the transplant bag. Mix during transfer.
Repeat with a second 60 ml syringe. The final volume is now approximately 170 ml (50 ml
diluted CBU and 120 ml dextran/albumin solution).
23. Close PC-3 and open PC-1 and PC-2. Pass the diluted CBU back and forth between the
transplant bag and the freezing bag in order to more completely wash all cells out of the
freezing bag and into the transplant bag. Close PC-4.
24. Seal tubing between PC-4 and Injection Port 1 (IP-1) and disconnect. Discard the spikes, luer
lock and connecting tubing.
25. Place the transplant bag and the waste bag in a centrifuge cup. Support the transplant
bag to prevent formation of creases during centrifugation. Make sure PC-5 and Screw Clamp 1 
(SC-1) are closed.
26. Centrifuge at 400 x g for 20 minutes at 10°C.
Note: You may now call the Transplant Unit and advise them that the transplant will be ready in 
about 30 minutes.
27. Place the centrifuged transplant bag in the plasma expressor. Open PC-5. Use SC-1 to
adjust the flow and very slowly transfer most of the supernatant to the waste bag. Leave
approximately 10-15 ml supernatant with the cells. Empty the tubing between the bags by
transferring air from the waste bag to the transplant bag. Close PC-5.
Note: If you detect passage of cells to the waste bag, return contents to the transplant bag,
re-suspend the cells and repeat the centrifugation.
28. Inspect the supernatant for escaped cells.
a. Centrifuge 10 ml of supernatant at 600 x g for 10 min.
b. Inspect the sediment for leukocytes in a microscope. If cells are found, count them and
calculate the total number of cells that would be lost. 
c. Decide whether the escaped cells should be recovered in a second centrifugation for
adding to the transplant cell dose. In that case, centrifuge the waste bag at 400 x g for 20
min. and transfer the supernatant to another waste bag (300 ml). Proceed following the
guidelines d + e.
d. If <75% of total viable nucleated cells are recovered, re-spin the supernatant in the waste bag 
to recover additional cells.
e. If ≥1x106 cells/kg of patient body weight are recovered, transport residual cells to the
patient’s room for reinfusion. If >1x105 but <1x106 cells/kg are recovered, cells should be
cryopreserved in 10% DMSO for future testing, storage or other uses. These cells can be
stored in a small vial or cryo bag at the discretion of the transplant center lab.
29. Seal the tubing between the bags close to the transplant bag. Disconnect the waste bag
with the supernatant.
30. Resuspend the cell pellet by slowly adding 25-50 ml of dextran-albumin solution in a 60
ml syringe through the IP-1. Mix cells and solution thoroughly during transfer.
31. Calculate the weight of the cells by subtracting the weight of an empty transplant bag.
32. Remove a small volume for cell count and viability determination. Samples for
bacteriology can be taken from the supernatant.
33. If cells are recovered from the first post-thaw supernatant, add 10 ml of dextran-albumin 
solution to the bag through IP-2. Resuspend the cells carefully.
Note: The volumes injected in step 30 may be modified if the Transplant Physician prefers

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injecting the patient with a smaller total volumen. In this case, resuspend the cell pellet to the 
desired volumen by injecting the desired volume of dextran-albumin solution.
34. Bring the transplant bag to the Transplant Unit even if the second centrifugation is being 
prepared; the second can be infused separately afterwards.
35. Prepare a report on the procedure. Note the condition of the bag and whether leaks or
cracks were detected. Include in the report:
- CBU ID
- Date of receipt of the CBU
- Storage conditions in your facility
- Date of thawing
- Cell count
- Viability of the cells recovered
Submit the report to the Transplant Unit and to StemCare. Alternatively or in addition fill out  
the Form QA/007E “Transplant Center Feedback Sheet” and return it to StemCare.
 

Protocol Version 1: 10 January 2019. Corrected 26.06.19; 17.06.21, Revised 14 October 2021             
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Clinical Study Protocol No: B-F011  
 
                           
 
                                                
 
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Annex 6 – Infusion protocol
Form Version 1 Printed from StemCare QMS 2019-01-22 Page 1 of 1
CBU Infusion Procedures
Principle
At the transplant center, the cryoprotected cord blood unit (CBU) is stored in the vapor phase of 
liquid nitrogen until the day of transplant, when it is thawed and washed in dextran / albumin, a 
process which increases cell viability and removes approximately 90% of the DMSO/dextran 
cryoprotectant. The washed cells are resuspended in dextran/albumin and transported to the 
patient’s bedside in a labelled transplant bag for infusion.
The unit is infused into the patient’s blood via the central venous catheter over 2-30 minutes.
Every effort should be made to be sure that all the cells in the transplant bag and IV tubing
are delivered to the patient, maintaining a closed system during the infusion.
Specimen
Thawed and washed cells from a cryopreserved cord blood unit (CBU) in a labelled
transplant bag.
Reagents
500 ml bag of Normal Saline
Supplies
Y-infusion set
IV extension tubing
Blood filter, 170-260 microns
Procedure
1. Verify that the patient is stable and has received premeds for transplant.
2. Verify that the transplant bag label matches patient identifiers via institutional 
procedures.
3. Examine transplant bag to be sure that cells are in solution and that large clumps are not
present in the bag. If clumps are present, prepare to use blood filter in infusion set-up.
4. Close all clamps on the tubing on the Y-infusion set and blood filter.
5. Spike one arm of the Y-infusion set into the bag of normal saline.
6. Connect extension tubing to the distal arm of the Y-infusion set.
7. Prime all tubing with normal saline.

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8. Spike the other arm of the Y-infusion set into the transplant bag.
9. Connect the distal end of the extension tubing to the patient’s central venous catheter.
10. Open the clamps between the transplant bag, the extension tubing, and the patient’s
central line and allow cells to infuse into the patient’s blood.
11. Rinse residual cells in the transplant bag and tubing. After all cells have dripped out of
the transplant bag, close the clamps on the extension tubing and Y-set connected to the
transplant bag. Open the clamps between the normal saline bag and transplant bag and
allow approximately 25 cc of saline to run into the transplant bag. Close the clamps 
between
the saline bag and transplant bag and open the clamps between the transplant bag and the
patient and infuse this saline to the patient. Repeat rinse x 1.
12. Monitor the patient’s vital signs before, during and after infusion per institution