Evaluation of Therapeutic Potential of Stromal Vascular Fraction (Autologous Adipose Derived Mesenchymal Stem Cell) Based Treatment for Chronic Kidney Disease
Phase: PHASE1
Conditions: Chronic Kidney Diseases
Sponsor: Bangladesh Laser & Cell Surgery Institute & Hospital



1 
 
RESEARCH PROTOCOL 
 
 
 
 
 
 
 
EVALUATION OF THERAPEUTIC POTENTIAL 
OF 
STROMAL VASCULAR FRACTION 
(AUTOLOGOUS ADIPOSE DERIVED 
MESENCHYMAL STEM CELL) BASED 
TREATMENT FOR CHRONIC KIDNEY DISEASE 
 
 
 
 
 
 
 
 
 
 
 

2 
 
TABLE OF CONTENT 
 
 
Topic 
Page No. 
List of Investigators 
3 
Place of Study 
4 
Sponsor 
4 
Study Description 
 
 
1. 
Title of the Study 
6 
2. 
Introduction 
6 
 
2.1 Chronic Kidney Disease 
6 
 
2.2 Stromal Vascular Fraction 
7 
 
2.3 Stem Cells 
7 
 
2.4 Advantages of Adipose Tissue-Derived Stem Cells 
8 
 
 
2.5 Kidney Disease and Mesenchymal Stem Cells 
9 
 
 
2.6 Clinical Trials Using MSC for Renal Repair 
9 
 
 
2.7 How Else SVF Help Tackle Kidney Diseases 
10 
 
 
2.8 Route of Delivery 
11 
 
 
2.9 Conclusion 
12 
 
3. 
Objectives of the Study 
13 
 
4. 
Methodology 
13 
 
5. 
Ethical Considerations 
24 
 
6. 
Resources 
24 
 
7. 
Procedure 
26 
Annexure 
 
 
A. 
References 
30 
 
B. 
Consent 
33 
 
C. 
Questionnaire – 1 
35 
 
 
Questionnaire – 2 
36 
 
D. 
Investigations – 1 
37 
 
 
Investigations – 2 
38 
 
 
Investigations – 3 
39 
 
 
 
 

3 
 
LIST OF INVESTIGATORES 
Principal Investigator(s):  
                 
 
 
Prof. DR. Md. Firoj Khan 
 
MBBS, MD (Nephrology) 
FRCP ( England), ISN Fellow ( Japan) 
Kidney Disease Specialist  
Consultant (Nephrology) 
             Bangladesh Laser & Cell Surgery Institute & Hospital,  
Dhaka, Bangladesh. 
Co-Investigator(s): 
1)   Dr. Mohammed Yakub Ali 
 
                                    MBBS, M.Phil, MSc, PhD. 
                                    Consultant (Laser Surgery) and  
                                    Director, Institute of Laser Surgery & Hospital and 
                                    Bangladesh LASER & Cell Surgery Institute & Hospital, 
                                    Dhaka. Bangladesh. 
                                   +88 01745490789  
 
 
 
                                    E-mail: myalibd@hotmail.com 
 
2)   Dr. Jahangir Md. Sarwar 
                                    MBBS, FCPS (Surgery) 
                                    Chief Consultant 
                                    Bangladesh Laser & Cell Surgery Institute & Hospital, 
                                    Dhaka, Bangladesh. 
                                    +88 01714044154 
                                    E-mail: jmsarwar2002@gmail.com 
 
 
 
 

4 
 
 
 
 
3)  Dr. Nibedita Nargis 
 MBBS, FCPS,MD 
 Senior Consultant (Anaesthesiology)                        
                                Bangladesh Laser & Cell Surgery Institute & Hospital, 
                                Dhaka, Bangladesh. 
                                +88 01712922266 
                                E-mail: nnargis255@gmail.com 
 
4) Dr. Mohammad Nazmul Kayes 
                                  MBBS, DA  
                                  Associate Professor (Anesthesiology) 
                                  Bangladesh Laser & Cell Surgery Institute & Hospital, 
                                  Dhaka. Bangladesh. 
                                  +88 01818281939 
                                  E-mail: nkayes.kiron@gmail.com 
5)   Dr. Mohammad Shahadat Hossain 
                                    MBBS. 
                                 Resident Surgeon 
                                 Bangladesh Laser & Cell Surgery Institute & Hospital,  
                                 Dhaka. Bangladesh. 
                                 +88 01714043555  
                                 E-mail: mshujjal@gmail.com 
 
6)   Dr. Afsana Sultana 
  MBBS 
  Medical Officer 
                                    Bangladesh Laser & Cell Surgery Institute & Hospital 
                                 Dhaka. Bangladesh. 
  +88 01737702646 
                                 E-mail: afsanasultana85@yahoo.com 
 

5 
 
PLACE OF STUDY 
 
Bangladesh Laser & Cell Surgery Institute & Hospital 
House No: 2, Road No: 2 
Sector: 2, Block : D 
Aftabnagar Housing, Badda, Dhaka – 1212 
Bangladesh 
Phone No: +88 02 55046715, +88 02 55046716 
E-mail: info@blcshospital.com 
Office E-mail: blcshospital@yahoo.com 
www.blcshospital.com 
 
 
SPONSOR 
Bangladesh Laser & Cell Surgery Institute & Hospital 
House No: 2, Road No: 2 
Sector: 2, Block : D 
Aftabnagar Housing, Badda, Dhaka – 1212 
Bangladesh 
 
 
 
 
 

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STUDY DESCRIPTION 
1. Title of the study 
Evaluation of Therapeutic Potential of Stromal Vascular Fraction (Autologous Adipose Derived 
Mesenchymal Stem Cell) Based Treatment for Chronic Kidney Disease 
2. Introduction 
2.1 Chronic Kidney Disease (CKD) 
CKD is a disease of alarmingly increasing prevalence (8 to 16%) associated with mortality [1]. 
CKD can progress towards end-stage renal disease (ESRD), requiring renal replacement therapy. 
ESRD currently accounts for 6.3% of the medicare spending in the United States, and is 
projected to increase by 85% by 2015 [2]. In a study conducted among rural population in 
Bangladesh overall CKD prevalence was found about 19% [3]. Furthermore, ESRD has a major 
impact on quality of life and life expectancy [4]. Therefore, it is very important to develop 
therapeutic interventions to prevent, alleviate, or decelerate progression of renal failure. 
Diabetes mellitus and hypertension represent major causes of CKD and initiation of dialysis [5]. 
In addition, glomerular diseases, malnutrition, infectious diseases, and acute kidney injury can 
progress to ESRD, contributing to the increased global burden of death [6]. Current treatment 
modalities often fail to target the major underlying contributors for progression of renal disease 
[7]. Management of CKD at present mostly aims at control of the predisposing factors and 
supplementation of kidneys homeostatic functions but not at the treatment of the diseased kidney 
itself. Again due to lack of adequate facilities or financial constraints people of a developing 
country like Bangladesh are unable to continue long term or lifelong dialysis. Chronic 
glomerular and tubule-interstitial fibrosis is a common pathway to ESRD, often associated with 
apoptosis, oxidative damage, fibrosis and microvascular rarefaction. Unfortunately, the 
regenerative potential of kidney is limited under chronic conditions and inefficient to prevent 
progressive glomerulosclerosis and tubule-interstitial fibrosis [8]. Treatment strategies that boost 
cellular regeneration might therefore offer good alternatives for patients with CKD.  

7 
 
In the western world as well as in Bangladesh, high blood pressure and type II diabetes are on 
the rise and are contributing to higher rates of kidney disease. Demand for kidney transplants is 
increasing. But there are not enough donor organs to meet this growing need. So, cell based 
therapies may offer an alternative solution. 
2.2 Stromal vascular fraction (SVF) 
SVF of adipose tissue is a rich source of pre-adipocytes, mesenchymal stem cells (MSC), 
endothelial progenitor cell, T cells, B cells, mast cells as well as adipose tissue macrophages 
[9,10]. SVF is a component of the lipo-aspirate obtained from liposuction of subcutaneous tissue. 
Lipo-aspirate contains a large population of stem cells called adipose derived stem cells 
(ADSCs), which share a number of similarities with bone marrow stem cells, including the 
capacity for multilineage differentiation . 
 
Figure 1: Stromal vascular fraction. 
2.3 Stem Cells 
A stem cell is a generic term referring to any unspecialized cell that is capable of long-term self-
renewal through cell division but that can be induced to differentiate into a specialized, 
functional cell. Stem cells are generally two types, embryonic stem cells and adult stem cells. 
Adult stem cells can be obtained from many differentiated tissues including, but not limited to, 
bone marrow, bone, fat, and muscle. Obtaining adult stem cells also does not raise any ethical 
concerns. For most studies, the adult stem cell in question is actually a mesenchymal stem cell 

8 
 
(MSC) or mesenchymal stromal cell. They are multipotent but not pluripotent, which means they 
can differentiate into some, or "multiple," but not all tissue types [11]. Stem cells that are 
harvested from the patient with the intention of administering them back to the same patient are 
termed autologous MSCs. MSC can also be isolated from the bone marrow (bmMSC), peripheral 
blood, connective tissue, skeletal muscle, dental pulp (dpMSC), umbilical cord wall (ucMSC), 
umbilical cord blood (cbMSC), amniotic fluid (afMSC) and all have been used in experimental 
settings to treat various types of renal diseases. An important feature of MSCs is their capacity to 
induce proliferation of renal glomerular and tubular cells, increasing cellular survival [12]. 
 
Figure 2: Sources of MSC used in experimental models of renal injury. Preclinical studies have 
shown that MSC used to treat renal diseases can be isolated from the following tissues: (A) tooth 
pulp, (B) kidney, (C) adipose tissue, (D) umbilical cord, (E) amniotic fluid, and (F) bone 
marrow. 
2.4 Advantages of adipose tissue-derived stem cells (ADSCs) 
ADSCs are somatic stem cell population contained in fat tissue and have been shown to possess 
stem cell properties such as trans-differentiation and self-renewal [13]. Similar to other types of 

9 
 
MSCs, ADSCs express multiple CD marker antigens (CD73+CD90+CD105+ CD34+/- CD11b- 
CD104b- CD19- CD31- CD45- SMA-) [14,15]. Additionally, utilizing ADSCs is advantageous 
in that large quantities of stem cells are easily isolated using minimally invasive surgical 
procedures [16]. 
ADSCs are vascular precursor cells. Many studies have shown that SVF contains progenitor cells 
that are able to differentiate into endothelial cells and participate in blood vessel formation [17]. 
Additionally, a recent study demonstrated that SVF cells expressing both pericyte and 
mesenchymal markers reside in a peri-endothelial location and stabilize endothelial networks 
[17] Another study showed that ADSCs transplanted into an ischemic renal cortex preferentially 
migrate toward micro vessels where they differentiate into vascular smooth muscle cells [18]. 
Some trials on kidney transplant recipients as well as the one on FSGS and 2 on CKD patients 
include in their protocol the utilization of adMSC. Adipose tissue is an important source of MSC, 
with a frequency 100 to 1000 times higher than bmMSC. They also seem to possess a higher 
potential for angiogenesis or vasculogenesis [19]. 
2.5 Kidney disease and mesenchymal stem cells 
A number of different types of cells from the bone marrow have been tested in animals and in 
clinical studies for potential use in kidney disease. Amongst all the cells under investigation, 
MSCs have shown the most promising results to date as they help kidney cells to grow, inhibit 
cell death and encouraging the kidney's own stem cells to repair kidney damage [20]. 
2.6 Clinical trials using mesenchymal stem cells for renal repair 
Few clinical trials have tested safety and efficacy of MSCs for renal disease. Reinders and 
colleagues studied safety and feasibility in six kidney allograft recipients who received two 
intravenous infusions of expanded autologous bone marrow-derived MSCs [21]. Importantly, 
delivery of autologous MSCs was not associated with adverse events, nor did it compromise 
graft survival. Several clinical trials are currently underway to evaluate the therapeutic potential 
of autologous and allogeneic MSCs for treatment of renal diseases [22] Administration of both 
bmMSC and adMSC has demonstrated significant reno-protective effects including reduction of 
intrarenal inflammatory infiltrate, decreased fibrosis, and glomerulosclerosis [12]  

10 
 
2.7 How else could stromal vascular fraction (autologous adipose derived MSC) help tackle 
kidney disease? 
MSCs possess unique immunomodulatory properties that ameliorate inflammation and immune 
responses, constituting a promising tool to facilitate renal repair. In recent years, experimental 
studies have uncovered the potential of MSCs to improve renal function in several models of 
CKD, and several clinical studies have indicated their safety and efficacy in CKD [22]. 
ADSCs could be incorporated into damaged tissues or organs which could give rise to new 
functional 
components 
and 
also 
exert 
potent 
anti-inflammatory, 
anti-fibrotic, 
or 
immunomodulation effects through paracrine or autocrine routes (via vascular endothelial 
growth factor, granulocyte/macrophage colony stimulating factor, stromal-derived factor-1alpha 
and hepatocyte growth factor) [23,24]. Interestingly, it is proposed that even apoptotic or dying 
ADSCs exhibit distinctive immunosuppressive properties [25]. ADSCs have been shown to 
possess stronger anti-inflammatory and immuno-modulating functions than bone marrow derived 
MSCs [26]. 
Villanueva et al. explored the effect of ADSCs on CKD by a single intravenous infusion of 
ADSCs on a nephrectomy induced CKD model of rats [27]. ADSC treatment was associated 
with reduced plasma creatinine, higher levels of epitheliogenic and angiogenic proteins, and 
improved renal function. Work by Hyun et al[28] illustrated the beneficial effects of ADSCs on 
improving renal function on a IgAN mouse model. Zhang et al.[29] found that repeated systemic 
administration of ADSCs attenuated proteinuria, glomerulus hypertrophy, and tubular interstitial 
injury in a DN rat model. 
Currently, several Clinical trials have been uploaded in the NIH database, all aim to test mainly 
the safety of using MSC and their efficacy in treating CKD. Two of them propose the use of 
autologous bmMSC and two adMSC. A study conducted in Tehran, Islamic Republic of Iran, 
which was designed to provide confirmation of Mesenchymal stem cell therapy in CKD. There 
18 months safety and efficacy of autologous MSC as a therapy for CKD total of 10 patients were 
conducted with I/V injection of high dose of 2x106 / kg of autologous MSC. Assessments were 
performed at 1,3,6,12 and 18 months after cell injections [30]. 

11 
 
Another study conducted in Birmingham, Alabama, Rochester, Minnesota, Jackson, Mississippi, 
USA where stem cell product called "Mesenchymal stem cell" grown from person's own fat 
tissue infused back in to the patient's own kidney and primary outcome measured after 3 months 
where renal tissue oxygenation increased and decrease in kidney inflammation was seen as 
secondary outcome [31]. 
2.8 Route of delivery 
Various routes for delivery of ADSCs, ADSC-induced cells, or ADSCs combined with 
compound materials have been developed for the treatment of different diseases or damaged 
tissue. These routes can be classified into two categories: systemic delivery through blood 
vessels (intravenous injection or intra-arterial injection) or local delivery directly into injured 
tissues or organs [32]. 
The route of MSC delivery may influence the cells' capacity to home and engraft the damaged 
tissue, and thereby their efficacy for renal repair. Commonly used experimental methods to 
deliver MSCs include systemic intravenous, intra-arterial, or intra-parenchymal delivery. In 
nonhuman primates the cells distribute broadly into the kidneys, skin, lung, thymus, and liver 
with estimated levels of engraftment ranging from 0.1 to 2.7% [33]. 
The route of MSC delivery, intravenous, intra-arterial, or intra-parenchymal, may affect their 
efficiency for kidney repair. When labeled MSC intravenously infused into baboons were 
observed for 9-21 months, estimated levels of engraftment in the kidney, lung, liver, thymus, and 
skin ranged from 0.1-2.7% [33] Indeed, the intravenous route lags in delivery efficiency, because 
MSC may initially be trapped in the lungs[34] Intra-arterial infusion of MSC was the most 
effective route to achieve immunomodulation in rat kidney transplantation, possibly by avoiding 
lodging in the pulmonary circulation, allowing MSC to home to the injured kidney [35]. 
An important feature of MSCs is their capacity to induce proliferation of renal glomerular and 
tubular cells, increasing cellular survival. By secreting proangiogenic and trophic factors, 
injected MSCs not only can enhance proliferation, but also can decrease apoptosis of tubular 
cells [36]. Several routes of administration (intra-parenchymal, sub-capsular, intravenous) have 

12 
 
been explored and all seem to be effective. Multiple, repeated injections of MSCs appear to be 
even more effective than single injections [37,38]. 
2.9 Conclusion 
Time has come to rethink about newer treatment approaches for increasing burden on 
haemodialysis and renal transplantation for CKD. MSCs have been shown to help avoid and 
reduce dialysis. It allows patients to work and continue life as productive citizens and also 
lessens dependency on family members by regaining independence. The financial benefits of 
eliminating dialysis and its consequences greatly outweigh the costs of stem cells. The long-term 
expense of dialysis is replaced by a short protocol of MSCs from SVF, which in all likelihood 
will eventually be a covered benefit of some insurance plans. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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3. Objectives of the Study 
 
3.1 General Objective 
To assess the outcome of stromal vascular fraction (autologous adipose derived mesenchymal 
stem cell) based treatment as a new, safe and minimal invasive modality for the treatment of 
chronic kidney disease. 
 
3.2 Specific Objectives: 
(i) To assess the safety of stromal vascular fraction (autologous adipose derived mesenchymal 
stem cell) in the treatment of CKD 
(ii) To determine the role of SVF in treating different stages of CKD  
(iii) To see if SVF can reverse or limit the progression of CKD  
 
4. Methodology 
The study will be conducted considering following methodological aspects. 
4.1 Study design  
Study Type:  
Interventional. 
Primary Purpose:  
Treatment 
Study Phase:  
Phase 1/Phase 2 
Interventional Study Model: Parallel Assignment  
Participants will be placed in either of the 2 groups, depending on 
the number of harvested total stem cell count. 
Those having a harvested total "Adipose Derived Stem Cell 
(ADSC)" count (in 5 ml SVF solution) between 1 x 10^6 to 2 x 
10^6 will be placed in "Group A". 

14 
 
Those having a harvested total "Adipose Derived Stem Cell 
(ADSC)" count (in 5 ml SVF solution) more than 2 x 10^6 will be 
placed in "Group B". 
Both groups will be studied for Phase I/ Phase II trial by the 
proposed outcome measures over the stipulated time frame. 
Those having a harvested total "Adipose Derived Stem Cell 
(ADSC)" count (in 5 ml SVF solution) less than 1 x 10^6 will be 
excluded from the study. 
Number of Arms:  
2 
Masking:  
Triple (Participant, Care Provider, Outcomes Assessor)  
Allocation:  
Non-Randomized 
Enrollment:  
31 [Anticipated] 
4.2 Arms and Interventions 
Arms: Participants having a harvested total "Adipose Derived Stem Cell (ADSC)" count (in 5 
ml SVF solution) between 1 x 10^6 to 2 x 10^6 will be placed in "Group A". 
Participants having a harvested total "Adipose Derived Stem Cell (ADSC)" count (in 5 
ml SVF solution) more than 2 x 10^6 will be placed in "Group B". 
Interventions : Both group A and Group B will be injected intravenously with harvested 5 ml of 
SVF containing Autologous Non Expanded ADSC and outcome will be observed over the period 
of 1(one) year. 
 
4.3 Study duration 
From April 2019 Onwards (Approximately Five years / Till completion of sample 
requirements with minimum one year of follow up). 
4.4 Study place 
Bangladesh Laser & Cell  Surgery Institute & Hospital. 
House No: 2, Road No: 2, Sector No: 2, Block : D 
Aftabnagar Housing, Badda, Dhaka – 1212. Bangladesh. 
 

15 
 
4.5 Study population: 
Patient of CKD who fulfill the selection criteria and admitted at the selected health 
facility for treatment.  
 
4.6 Sample size determination: 
α (two-tailed) =   Threshold probability for rejecting the null hypothesis. Type I error rate.  
β =    Probability of failing to reject the null hypothesis under the alternative hypothesis. Type II 
error rate.  
E =   Effect size  
S(Δ) =   Standard Deviation of the change in the outcome.  
Here, α = 0.05 
β = 0.20 
E = 0.5 
S(Δ) = 1.0 
The standard normal deviate for α = Zα = 1.960 
The standard normal deviate for β = Zβ = 0.842 
A = 1.000 
B = (Zα+Zβ)2 = 7.849 
C = (E/S(Δ))2 = 0.250 
Sample size N = AB/C = 31 (approx.) 
 

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4.7 Sampling technique & Recruitment of study subjects 
Participants of the study will be recruited from the patients with CKD attending the Bangladesh 
Laser & Cell Surgery Institute & Hospital considering the eligibility criteria. The recruitment 
will be continued till the desired sample is drawn.  
4.8 Eligibility criteria 
4.8.1 Inclusion criteria 
A patient is eligible for the study if all of the followings apply: 
a) Aged 18-80 years (inclusive) 
b) With chronic kidney disease (CKD)stage 3 to 5 (eGFR 60 to 0 mL/min/1.73m2 
(inclusive)) Note : eGFR = estimated glomerular filtration rate 
c) Having provided informed written consent. 
4.8.2 Exclusion criteria 
Any patient meeting any of the exclusion criteria will be excluded from study 
participation. 
1) Known hypersensitivity to any component used in the study. 
2) With inadequate hematologic function with: absolute neutrophil count (ANC) 
<1,500/μL OR platelets < 100,000/μL OR Hemoglobin < 8 g/dL 
3) With impaired hepatic function with: serum bilirubin, aspartate aminotransferase 
(AST), alanine aminotransferase (ALT) or alkaline phosphatase (AKP), prothrombin 
time above and normal reference and serum albumin below normal reference range. 
4) With hemoglobin A1c (HbA1c) > 8.0% 
5) With serious prior or ongoing medical conditions (e.g. concomitant illness such as 
cardiovascular (e.g. New York Heart Association grade III or IV), hepatic e.g. Child-
Pugh Class C), psychiatric condition, alcoholism, drug abuse), medical history, 
physical findings, ECG findings, or laboratory abnormality that in the investigators' 
opinion could interfere with the results of the trial or adversely effect the safety of the 
patient 

17 
 
6) Pregnant or lactating women or premenopausal with childbearing potential but not 
taking reliable contraceptive method(s) during the study period 
7) With known history of human immunodeficiency virus (HIV) infection or any type of 
hepatitis 
8) Judged to be not applicable to this study by investigator such as difficulty of follow-
up observation 
9) With any other serious diseases/medical history considered by the investigator not in 
the condition to enter the trial 
10) Known or suspected abuse of alcohol or narcotics 
11) With known history of cancer within past 5 years 
12) With any autoimmune disease 
13) With congenital kidney disease 
14) With precancerous condition or with raised tumour markers like Alpha feto protein, 
Carcino embryonic antigen (CEA), C.A 19.9, C.A 125, Serum PSA above normal 
reference range. 
15) Participants having a harvested total “Adipose Derived Stem Cell (ADSC)” count (in 
5 ml SVF solution) less than 1 x 10^6 will be excluded from the study. 
 
4.9 Treatment allocation  
All subjects would be submitted for Stromal Vascular Fraction (Autologous Adipose 
Derived Mesenchymal Stem Cell) Based Treatment for Chronic Kidney Disease 
4.10 
Pre-transplant and Post-Transplant Investigations  
Investigations will be done according to the Annexure – D 
4.11 
Outcome measures 
4.11.1 Primary Outcome Measure  
a) Incidence of minor adverse events (MAEs) , serious adverse events (SAEs) which may be 
immediate, early or late - for Phase I   [Time Frame: Week 48]  
Minor adverse events (MAEs): 
i) Pain from lipo-suction > 7 days (Early) 

18 
 
ii) Fever > 7 days (Early) 
iii) Subcutaneous hematoma / abscess formation (Early) 
iv) Allergic reaction (Immediate) 
Serious adverse events (SAEs) 
i) 
Anaphylaxis (Immediate) 
ii) 
Pulmonary embolism or infarction (Immediate) 
iii) 
Outset of any neoplastic change (Late) 
iv) 
Outset of new Cardiovascular events (Late) 
v) 
Outset of new Cerebrovascular or neurological events (Late) 
vi) 
Reactivation of treated tuberculosis (Late) 
b) Change from baseline to 24 week visit in estimated glomerular filtration rate (eGFR) and 
split renal function in all patients - for Phase II   [Time Frame: Weeks 0, 24]  
eGFR with split renal function will be evaluated using DTPA Renogram. 
c) Change from baseline to 24 week visit in estimated glomerular filtration rate (eGFR) with 
serum creatinine level in patients with CKD 4 and below - for Phase II   [Time Frame: 
Weeks 0, 2, 4, 12, 24]  
eGFR will be calculated by Serum Creatinine level using MRDR formula during all 
visits. 
d) Change from baseline to 24 week visit in need for dialysis in patients with CKD 5 - for 
phase II   [Time Frame: Weeks 0, 2, 4, 12, 24]  
Need for dialysis is described as 
i)       No dialysis needed - Score 0 
ii) 
Randomly (more than 6 days interval) - Score 1 
iii) 
At 6 (six) days interval / Once weekly - Score 2 
iv) 
At 5 (five) days interval - Score 3 
v) 
At 4 (four) days interval - Score 4 

19 
 
vi) 
At 3 (three) days interval / 2 times a week - Score 5 
vii) 
At 2 (two) days interval - Score 6 
viii) 
At 1 (one) day interval / every alternate day./ 3 times a week - Score 7 
4.11.2 Secondary Outcome Measures  
a) Change from baseline to all post-treatment visits in body weight   [Time Frame: Weeks 0, 
2, 4, 12, 24, 36, 48]  
Weight in Kg will be recorded for each patient during each follow up 
b) Change from baseline to all post-treatment visits in Blood-pressure   [Time Frame: 
Weeks 0, 2, 4, 12, 24, 36, 48]  
Blood pressure will be measured in each patient during each follow up 
c) Change from baseline to all post-treatment visits in S.creatinine   [Time Frame: Weeks 0, 
2, 4, 12, 24, 36, 48]  
a. S. Creatinine level will be measured during each follow up. 
b. In case of patients having dialysis Pre and Post dialysis S. Creatinine levels will 
be measured at or near follow up dates. 
d) Change from baseline to all post-treatment visits in blood urea nitrogen (BUN)   [Time 
Frame: Weeks 0, 2, 4, 12, 24, 36, 48]  
Blood Urea Nitrogen will be measured in all patients during each follow up. 
e) Change from baseline to all post-treatment visits in Hemoglobin level   [Time Frame: 
Weeks 0, 2, 4, 12, 24, 36, 48]  
a. Hemoglobin level will be measured in gm/dl and percentage 
b. Need for blood transfusion will be recorded 
c. Need for erythropoietin will be recorded 
f) Change from baseline to all post-treatment visits in urine microalbumin-to-creatinine 
ratio (UMCR)   [Time Frame: Weeks 0, 2, 4, 12, 24, 36, 48]  

20 
 
Urinary Microalbumin and creatinine will be measured in each patient during each follow 
up 
g) Change from baseline to all post-treatment visits in hemoglobin A1c   [Time Frame: 
Weeks 0, 2, 4, 12, 24, 36, 48]  
HbA1C will be measured in each patient during each follow up 
h) Change from baseline to all post-treatment visits in random blood sugar (RBS)   [Time 
Frame: Weeks 0, 2, 4, 12, 24, 36, 48]  
RBS will be measured in each patient during each follow up 
i) Percentage of patients with hypoglycemia (defined as blood glucose < 55 mg/dL or 3.0 
mmol/L) at all post-treatment visits   [Time Frame: Weeks 0, 2, 4, 12, 24, 36, 48]  
Patients will be asked if he or she had experienced any episode of hypoglycemia with 
clinical features of altered level of consciousness, sweating, nausea or vomiting with 
blood glucose < 55 mg/dL or 3.0 mmol/L 
j) Change from baseline to all post-treatment visits in eGFR   [Time Frame: Weeks 0, 2, 4, 
12, 24, 36, 48]  
eGFR will calculated using MDRD formula for each patient during each follow up 
k) Change from baseline to all post-treatment visits in Anti-Hypertensive medication if there 
is any.   [Time Frame: Weeks 0, 2, 4, 12, 24, 36, 48]  
All anti hypertensive medicines with their doses will be recorded including any changes 
in each patient during each follow up 
l) Change from baseline to all post-treatment visits in Hypoglycemic agent if there is 
any.   [Time Frame: Weeks 0, 2, 4, 12, 24, 36, 48]  

21 
 
All hypoglycemic agents including their doses with any changes will be recorded for all 
diabetic patients during each follow up 
m) Change from baseline to post-treatment visits in urine total protein-creatinine ratio 
(UPCR)   [Time Frame: Weeks 0, 24, 48]  
Urinary total protein and Creatinine ratio will be done in each patient during defined 
visit. 
n) Change from baseline to post-treatment level of serum Alpha Feto Protein   [Time Frame: 
Weeks 0, 24, 48]  
Serum Alpha Feto protein will be measured as a tumour marker for Hepato-cellular 
carcinoma and also Tumour of Testis and Ovary. 
o) Change from baseline to post-treatment level of serum CEA level   [Time Frame: Weeks 
0, 24, 48]  
Serum CEA level will be measured as a tumour marker for Colo-rectal Carcinoma and 
also for Cancer of Stomach, pancreas, breast, lungs, thyroid and ovary. 
p) Change from baseline to post-treatment level of serum CA 19.9 level   [Time Frame: 
Weeks 0, 24, 48]  
Serum C.A 19.9 level will be measured as a tumour marker for Pancreatic Carcinoma 
q) Change from baseline to post-treatment level LDH level   [Time Frame: Weeks 0, 24, 48]  
Serum LDH level will be measured as tumour marker for Lymphoma 
r) Change from baseline to post-treatment level of Beta 2 Microglobulin level   [Time 
Frame: Weeks 0, 24,48]  
Serum Beta 2 Microglobulin level will be measured as a prognostic tool, as CKD patients 
invariably has a raised serum level. 

22 
 
s) Change from baseline to post-treatment level of serum CA 125 level (in case of female 
patients)   [Time Frame: Weeks 0, 24, 48]  
Serum C.A 125 level will be measured as a tumour marker for Ovarian Cancer 
t) Change from baseline to post-treatment level of PSA level (in case of male 
patients)   [Time Frame: Weeks 0, 24,48]  
Serum PSA level will be measured as a tumour marker for Prostatic Cancer. 
4.12 Research instrument  
For collection of primary data about patient a semi-structured questionnaire will be developed 
based on research objective. Pre-testing of the questionnaire will be done on other patients 
admitted at the same facility.  After the pretesting, amendment of the items and question will be 
done based on study finding. In the final questionnaire both structured and open questions will be 
kept.  
A check list will be prepared to compile the data from hospital records, treatment records, 
outcome of treatment and laboratory investigation reports.  
4.13 Data collection procedure: 
Current study involves collection of both primary and secondary data. Primary data will be 
collected by face to face interview of the patients or patient’s attendant by the researcher at 
health facility during the period of hospital stay, upon their consent and convenience. Socio 
demographic and personal information will be recorded from patient through interview, with a 
semi structured pre-tested questionnaire.  Information regarding risk factors and risk behavior 
will be inquired taking effort to minimize the recall bias.  
Secondary data about present state and diagnosis will be collected from hospital record as well as 
from treatment sheet. For acquiring secondary data a structured checklist will be used. 
 
 

23 
 
4.14 Data processing 
Data processing will include data cleaning and quality control check, editing of data, coding of 
data and data entry into computer. The edited data will then be entered on to the template of 
SPSS® 16 and STATA® 10/IC.  
4.15 Data analysis  
The edited data will then be entered on to the template of SPSS® 16 and STATA® 10/IC. 
For Back ground variables and socio-demographic data descriptive statistics and relative 
frequency (percentage) will be generated. 
Through univariate analysis the base line characteristics and treatment outcome will be 
compared. The effect of treatment will be identified through Multivariate analysis after adjusting 
for possible confounders. Relative risk with 95% CI will be generated through binary logistic 
regression adjusting for all possible confounders.   
4.16 Data Presentation 
Data will be presented in the form of table and graphs. Descriptive statistics will be presented 
with frequency table.  Association will be illustrated with cross tables and test statistics will be 
added in the foot note of the table. Bar and pie charts will be generated to illustrate descriptive 
statistics. 
 
 
 
 
 
 

24 
 
5 
Ethical considerations 
Approval has already been taken from 
National Research Ethics Committee, 
Bangladesh Medical Research Council (BMRC) 
Phone: +88029848396   Email: info@bmrcbd.org 
BMRC Bhaban, Mohakhali, Dhaka 1212, Bangladesh. 
 
Approval Number: 076 (1) 25 10 2017  
Informed written consent (Annexure - ) will be taken from the participant after explaining all 
the facts, potential dangers to the subjects in case of primary data collection. The study involves 
collection of non-sensitive Socio-demographic data (primary data), compilation of hospital and 
treatment records (secondary data) to be conducted researcher under supervision of relevant 
specialists along with few diagnostic test. The participants will be assured that the information 
acquired will be used for academic purpose. They will be assured of confidentiality, and for the 
purpose of data analysis no individual data were reported rather de identified data will be 
preceded for analysis.  
6 
Resources 
6.1 Space and Budget 
a) Principal and other investigators have their respective offices in the premises of the 
Bangladesh Laser & Cell Surgery Institute & Hospital, Dhaka, Bangladesh. 
b) “Bangladesh Laser & Cell Surgery Institute & Hospital” Dhaka, Bangladesh has 
adequate facilities to carry out the procedure in its operation theater. 
c) “Bangladesh Laser & Cell Surgery Institute & Hospital” Dhaka, Bangladesh has 
adequate facility to carry out post-operative management of the subjects of the current 
study. 

25 
 
d) “Bangladesh Laser & Cell Surgery Institute & Hospital” Dhaka, Bangladesh has 
proposed the budget for their operative and post- operative cost and the cost of the 
hospital stay. 
e) “Bangladesh Laser & Cell Surgery Institute & Hospital, Dhaka, Bangladesh”. has 
proposed the budget for data collection, data processing and rounding up the study over 
the expected time period. 
6.2 Facilities 
(a) Lab facilities (all modern equipments and Lab materials are available) 
(b) Man power facilities (mentioned investigators, Co- investigators, Anesthesiologist, 
Medical Officer, Nurse, Ward Boy) 
6.3 Materials 
a) Study population According to sample size calculation 31 patients with 
established CKD  will be recruited in the study over the estimated study period. 
b) Equipment “UNISTATIONTM” a specially designed medical device to satisfy 
diverse demands for various autologous cell therapies.  
UNISTATION is a specially designed medical device to satisfy diverse demands for various 
autologous cell therapies with only one device. PRP, PRF isolation, and fat purification functions 
have been added to UNISTATIONTM. 

26 
 
7 
Procedure 
After harvesting the abdominal adipose tissue via liposuction SVF (stromal vascular fluid) are 
collected. Subsequently these are centrifuged in UNISTATIONTM, a centrifuge machine 
specifically manufactured for this purpose. Later it is mixed with collagenase solution and fat is 
separated. Stem cells containing SVF is collected after centrifugation. SVF is neutralized and 
washed with normal saline. Then it is washed with patient’s own serum and normal saline. A 
sample is collected for cell counting by Luna automated cell counter. The final SVF is ready 
for subcutaneous or IV injection if 80-90% of cell remain viable, active and mobile. 
As it is a closed door automatic procedure within a same machine, so after collection of 40ml of 
sub-cutaneous fat, all procedures are automated. So there is no chance of contamination. Steps 
are mentioned below. 
1st Step:  Fat washing(A1) : 
(1) To aspirate 40 ml fat and to take 40 ml water for balance in 2 UNIKITs. 
(2) To put them is UNISTATIONTM Centrifuge. 
(3) To touch SVF button once (Display shows A1) 
(4) To touch start button once A1 is shown. 
(5) After centrifugation, to remove RBC layer from the bottom. 
(6) To collect 20ml pure fat after removing RBC layer in another UNIKIT. 
 
 
 
 
 
 
 

27 
 
2nd Step: Collagenase Digestion(A2) : 
(1) To prepare 20ml of 0.1% collagenase Solution. 
(2) To transfer the Collagenase solution in to the pure fat. 
(3) To place shaking plate and put the pure fat with collagenase on if and close the door 
(4) To touch SVF button once again (Display shows A2) 
(5) To touch Start button once A2 is shown. 
(6) To wait for 30 min for shaking incubation and take the fat out after finishing shaking 
incubation. 
 
 
 
 
 
 
 
 
 
3rd Step : SVF Collection(A3): 
(1) To put the collagen digested fat in UNISATIONTM Centrifuge. 
(2) To touch SVF button once again (Display shows A3) 
(3) To touch Start button once A3 is shown. 
(4) After centrifugation, to take out UNIKIT very slowly so that layers should not be 
mixed. (Stem cell are at the bottom in 5ml SVF) 
(5) Before removing cap to put the hand piece upwards so that stem cells should not be 
kept in the cap.  

28 
 
(6) To transfer 5ml SVF at the bottom into a 5ml or 10ml syringe and transfer it again 
into another UNIKIT. 
4th Step: Neutralization and Washing of SVF(A4) : 
(1) To transfer 5ml PPP into the 5ml SVF. 
(2) To transfer 30ml Normal saline into the 10ml of SVF and PPP. 
(3) To put it in UNISTATIONTM Centrifuge. 
(4) To touch SVF button once again (Display shows A4) 
(5) To touch Start button if A4 is shown. 
(6) After centrifugation. To transfer 5ml SVF at the bottom into 5ml or 10ml syringe. 
 
 
 
 
 
 

29 
 
 
Final Step: Preparation for Injection 
 
To transfuse the final 5 ml of SVF to the patient through intravenous route. Before that to 
collect few drops of SVF sample for cell counting by Luna Stem automated fluorescent cell 
counter.  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

30 
 
ANNEXURE - A 
References 
1.  
Jha V, Garcia-Garcia G, Iseki K, Li Z, Naicker S, Plattner B, Saran R, Wang AY, Yang CW: 
Chronic kidney disease: global dimension and perspectives. Lancet 2013, 382:260–272. 
2.  
Gilbertson DT, Liu J, Xue JL, Louis TA, Solid CA, Ebben JP, Collins AJ: Projecting the number of 
patients with end-stage renal disease in the United States to the year 2015. J Am Soc Nephrol 2005, 
16:3736–3741. 
3.  
Prevalence of Chronic Kidney Disease (CKD) and Identification of Associated isk Factors among 
Rulral Population by Mass Screening. Hasan MJ , Kashem MA,, Rahman MH, Quddhush R , 
Rahman M, Sharmeen A, Islam N. CBMJ 2012;1(1):20-26. 
4.  
Wyld M, Morton RL, Hayen A, Howard K, Webster AC: A systematic review and meta-analysis of 
utility-based quality of life in chronic kidney disease treatments. PLoS Med 2012, 9:e1001307. 
5.  
Collins AJ, Foley RN, Chavers B, Gilbertson D, Herzog C, Johansen K, et al. United States renal 
data system 2011 annual data report: atlas of chronic kidney disease & end-stage renal disease in 
the United States. Am J Kidney Dis 2012, 59:A7. e1-e420. 
6.  
Perico N, Remuzzi G: Chronic kidney disease: a research and public health priority. Nephrol Dial 
Transplant 2012;27:19–26. 
7.  
Rivera JA, O'Hare AM, Harper GM: Update on the management of chronic kidney disease. Am 
Fam Physician 2012;86:749–754. 
8.  
Benigni A, Morigi M, Remuzzi G: Kidney regeneration. Lancet 2010;375:1310–1317. 
9.  
Riordan, NH; Ichim, TE; Min, WP; Wang, H; Solano, F; Lara, F; Alfaro, M; Rodriguez, JP; 
Harman, RJ; Patel, AN; Murphy, MP; Lee, RR; Minev, B. "Non-expanded adipose stromal 
vascular fraction cell therapy for multiple sclerosis.". Journal of translational medicine. 2009;7:29.  
10.  Schipper HS, Prakken B, Kalkhoven E, Boes M. Adipose tissue-resident immune cells: key players 
in immunometabolism. Trends Endocrinol Metab 2012; 23:407-15. 
11.  Reinders, ME, Fibbe, WE, Rabelink, TJ. Multipotent mesenchymal stromal cell therapy in renal 
disease and kidney transplantation. Nephrol Dial Transplant 2010;25:17–24. 
12.  Peired AJ, Sisti A, Romagnani P. Mesenchymal Stem Cell-Based Therapy for Kidney Disease: A 
Review 
of 
Clinical 
Evidence. 
Stem 
Cells 
International 
2016. 
http://dx.doi.org/10.1155/2016/4798639. 
13.  Kokai LE, Rubin JP, Marra KG (2005) The potential of adipose-derived adult stem cells as a 
source 
of 
neuronal 
progenitor 
cells. 
Plast 
Reconstr 
Surg 
116:1453-1460. 
doi: 

31 
 
10.1097/01.prs.0000182570.62814.e3. 
14.  Lin CS, Xin Z, Deng C, Ning H, Lin G, et al. (2010) Defining adipose tissue derived stem cells in 
tissue and in culture. Histol Histopathol 25: 807-815. doi: 20376787. 
15.  Lin F (2012) Adipose tissue-derived mesenchymal stem cells: a fat chance of curing kidney 
disease?. Kidney Int 82: 731-733. doi: 10.1038/ki.2012.158.PMID: 22975993 
16.  Huixi Li1, 2, Guiting Lin1*, and Tom F Lue1 Potential application of adipose tissue-derived stem 
cells for urological disease. Bladder 2014;1(1). DOI: 10.14440/bladder.2014.23 
17.  Stashower M, Smith K, Williams J, Skelton H. Stromal progenitor cells present within liposuction 
and reduction abdominoplasty fat for autologous transfer to aged skin. Dermatol Surg 1999;25:945-
949. 
18.  Halvorsen YC, Wilkison WO, Gimble JM. Adipose-derived stromal cells- 
-their utility and potential in bone formation. Int J Obes Relat Metab Disord 2000;24 Suppl 4:41-
44. 
19.  Moseley TA, M. Zhu, and M. H. Hedrick, “Adipose-derived stem and progenitor cells as fillers in 
plastic and reconstructive surgery,” Plastic and Reconstructive Surgery 2006;118(3):121S–128S. 
20.  Kidney disease: how could stem cells help?.  http://www.eurostemcell.org /factsheet/kidney-
disease-how-could-stem-cells-help 
21.  Reinders ME, de Fijter JW, Roelofs H, Bajema IM, de Vries DK, Schaapherder AF, et al. 
Autologous bone marrow-derived mesenchymal stromal cells for the treatment of allograft 
rejection after renal transplantation: results of a phase I study. Stem Cells Transl Med 2013;2:107–
111. 
22.  Eirin A, Lerman LO. Mesenchymal stem cell treatment for chronic renal failure. Eirin and Lerman 
Stem Cell Research & Therapy 2014;5:83. 
23.  Rehman J, Traktuev D, Li J, Merfeld-Clauss S, Temm-Grove CJ, et al. Secretion of angiogenic and 
antiapoptotic factors by human adipose stromal 
cells. Circulation 2004;109:1292-1298. 
24.  Meliga E, Strem BM, Duckers HJ, Serruys PW. Adipose-derived cells. Cell Transplant 
2007;16:963-970. 
25.  Thum T, Bauersachs J, Poole-Wilson PA, Volk H, Anker SD. The dying 
stem cell hypothesis: immune modulation as a novel mechanism for progenitor 
cell therapy in cardiac muscle. J Am Coll Cardiol 2005;46:1799-1802. 
26.  Banas A, Teratani T, Yamamoto Y, Tokuhara M, Takeshita F, et al. IFATS collection: in vivo 
therapeutic potential of human adipose tissue mesenchymal stem cells after transplantation into 
mice with liver injury. Stem Cells 2008;26:2705-2712. 

32 
 
27.  Villanueva S, Carreño JE, Salazar L, Vergara C, Strodthoff R, et al. Human mesenchymal stem 
cells derived from adipose tissue reduce functional and tissue damage in a rat model of chronic 
renal failure. Clin Sci (Lond) 2013;125:199-210. 
28.  Quimby JM, Webb TL, Habenicht LM, Dow SW. Safety and efficacy of intravenous infusion of 
allogeneic cryopreserved mesenchymal stem cells for treatment of chronic kidney disease in cats: 
results of three sequential pilot studies. Stem Cell Res Ther 2013;4:48. 
29.  Zhang L, Li K, Liu X, Li D, Luo C, et al. Repeated systemic administration of human adipose-
derived stem cells attenuates overt diabetic nephropathy in rats. Stem Cells Dev 2013;22:3074-
3086. 
30.  D.D. Papazova, N.R. osterhuis, H. Gremmels, A. van Koppen, J.A.Joles, and M.C. Verhaar,"Cell-
based therapies for experimental chromic lidney disease: a systematic review and meta-analysis,” 
Disease Models and Meechanisms, vol.8, no.3 pp. 281-293, 2015. 
31.  L.Malaga-Dieguez, D. Bouhassira, D.Gipson, and H. Tracht-man,̒̒̒̒̒̒̒̒ Novel therapies for FSGS: 
preclinical and clinical studies,” Advance in chronic Kidney Disease, vol. 22, No. 2, pp.el-e6, 
2015. 
32.  You D, Jang MJ, Lee J, Suh N, Jeong IG, et al. Comparative analysis of periprostatic implantation 
and intracavernosal injection of human adipose tissue-derived stem cells for erectile function 
recovery in a rat model of cavernous nerve injury. Prostate 2012;73:278-286. doi: 
10.1002/pros.22567. 
33.  Devine SM, Cobbs C, Jennings M, Bartholomew A, Hoffman R: Mesenchymal stem cells 
distribute to a wide range of tissues following systemic infusion into nonhuman primates. Blood 
2003, 101:2999–3001. 
34.  Fischer UM, Harting MT, Jimenez F, et al. Pulmonary passage is a major obstacle for intravenous 
stem cell delivery: the pulmonary first-pass effect. Stem Cells Dev. 2009; 18:683–692. 
35.  Zonta S, De Martino M, Bedino G, et al. Which is the most suitable and effective route of 
administration for mesenchymal stem cell-based immunomodulation therapy in experimental 
kidney transplantation: endovenous or arterial? Transplant Proc. 2010; 42:1336–1340. 
36.  Bi B, Schmitt R, Israilova M, Nishio H, Cantley LG: Stromal cells protect against acute tubular 
injury via an endocrine effect. J Am Soc Nephrol 2007, 18:2486–2496. 
37.  Lee, SR, Lee, SH, Moon, JY, et al. Repeated administration of bone marrow-derived mesenchymal 
stem cells improved the protective effects on a remnant kidney model. Ren Fail 2010;32:840–848. 
38.  Semedo, P, Correa-Costa, M, Antonio Cenedeze, M, et al. Mesenchymal stem cells attenuate renal 
fibrosis through immune modulation and remodeling properties in a rat remnant kidney model. 
Stem Cells 2009;27:3063–3073. 
 

33 
 
ANNEXURE – B 
CONSENT 
Being fully conscious and oriented I, the undersigned willingly giving my consent  
1) To enroll myself as a subject of this new research oriented treatment protocol. I have 
been explained the pros and cons of this study including the already existing alternative 
treatment modalities for my disease conditions, the complications that may occur, the possible 
outcome of this treatment etc. I will not hold anybody responsible for any complication that may 
arise during the course of treatment or if the procedure ends up in failure. Rather I will comply 
with the salvage procedure that may be taken to manage the complications or in case of failure. 
2) To provide detail information regarding my disease condition and wellbeing without 
concealing anything particularly that of the co morbid disease conditions. 
3) To participate in the follow up sessions as mentioned in the treatment protocol and 
extend my full co-operation during the follow up proceedings. I must not conceal or exaggerate 
any information that may actively or passively influence the result of the research. 
 
 
 
 
 
 
 
 
 
       Please tick as appropriate 
1) Do you have complete idea about the type, ultimate goal 
and methodology of this treatment protocol? 
2) Are you aware of the existing treatment facility for your disease? 
3) Are you aware that you will not have any additional risk than that 
of the conventional treatment options ? 
4) Have you decided intentionally to take this treatment, despite knowing the 
possibility of poor / no result of the treatment?  
5) Do you think this research will violet your human rights? 
Yes 
No 
Yes 
No 
Yes 
No 
Yes 
No 
Yes 
No 

34 
 
6) Do you have confidence on the confidentiality of your records?  
7) Do you consent for your medical records to be used for further medical  
Research? 
 
 
______________________               _____________________            _____________________ 
(Signature of the Physician)  
(Signature of the Witness) 
          Signature of the Patient 
Name:  
 
 
 
Name:  
 
 
          Name: 
Address: 
 
 
 
Address: 
 
 
          Address: 
 
Yes 
No 
Yes 
No 

ANNEXURE - C
QUESTIONNAIRE - 1
(CLINICAL ASSESSMENT - DURING ALL VISITS)
PARTICULARS OF THE PATIENT:
CODE NO:
NAME: 
AGE:
SEX:
M  /  F
HOSPITAL REG. NO:
ADDRESS:
PHONE:
CONTACT PERSON:
PHONE:
DIAGNOSIS (Put Tick Mark Beside The Appropriate Selection):
1) CKD:
Stage 3A
Stage 3B
Stage 4
Stage 5
          a) GN
Need for dialysis
Score
          b) IgA Nephropathy
a) No dialysis yet
0
          c) FSGS
b) Occasional (> 6 days interval)
1
          d) Interstitial Nephritis
c) At 6 (six) days interval / Once weekly
2
          e) No Biopsy done
d) At 5 (five) days interval 
3
e) At 4 (four) days interval
4
f) At 3 (three) days interval / 2 times a week 
5
g) At 2 (two) days interval
6
h) At 1 (one) day interval / 3 times a week - 
7
2) HYPERTENSION
3) DM
4) Hypothyroidism
5) HYPERPARATHYROIDISM
6) IHD < NyHA Gr III or IV 
7) CLD < Child Pugh C
8) CVD
9) OTHERS (if any)
SYMPTOMS (Put Tick Mark Beside The Appropriate Selection):
Urine output:     
Scanty / Reduced / Normal
General Welbeing
Better/Good/As usual/Bad/Worse
Anorexia:
Yes / No
Convulsion:
Yes / No
Nausea:
Yes / No
Loss of Consciousness:
Yes / No
Vomiting:
Yes / No
Muscle spasm:
Yes / No
Hiccough:
Yes / No
Pain
Yes / No
Weakness:
Yes / No
Fever
Yes / No
Weight loss:
Yes / No
Headache:
Yes / No
Swelling of body:
Yes / No
Respiratory distress:
Yes / No
Vertigo:
Yes / No
Chest pain:
Yes / No
Constipation:
Yes / No
Abdominal Pain:
Yes / No
Loose motion:
Yes / No
Others (if any):
SIGNS:
Body weight:
Blood pressure:
Oedema:
Pulse:
Respiration:
Dehydration:
Temperature:
Others (if any):
35

ANNEXURE - C
QUESTIONNAIRE - 2
(CONCOMITANT TREATMENT - DURING ALL VISITS)
PARTICULARS OF THE PATIENT:
CODE NO:
NAME: 
AGE:
SEX:
REG. NO:
TYPE OF MEDICINE
NAME OF MEDICINE
DOSE
CHANGES IF ANY
ANTIHYPERTENSIVE
HYPOGLYCAEMIC
ANTIBIOTICS
ANALGESICS
ANTIULCERANT
VITAMINS & MINERALS 
OTHERS
36

ANNEXURE - D
(INVESTIGATIONS - 1ST VISIT - 0 WEEK)
PARTICULARS OF THE PATIENT:
CODE NO:
NAME: 
AGE:
SEX:
M / F
REG. NO:
INVESTIGATIONS for Outcome Measures:
INVESTIGATIONS for Fitness & Maintainence:
DTPA Renogram
Total GFR
BIOCHEMICAL:
Right
S.URIC ACID
Left
S. Vit D3 
S. Calcium
CBC/ESR
S. Phosphate
Hb (gm/dl)
ESR (mm in 1st hour)
S. ELECTROLYTES:
PLATELET COUNT (K/ul)
SODIUM
WBC 
POTASSIUM
Neutrophil (%)
CHLORIDE
Lymphocyte (%)
TCO2
Monocyte (%)
ANION GAP
Eosinophil (%)
Basophil (%)
HORMONES
S. PTH
BIOCHEMICAL:
S. TSH
RBS
FT4
HbA1C
S. Creatinine
LIVER PROFILE
e GFR (ml/min/1.73 m2)
HBsAg
BUN
Anti HCV
Urine Microalbumin-Creatinine Ratio (UMCR)
ALT 
Urinary Total Protein-Creatinine ratio(UPCR)
S. Albumin
Prothrombin Time
TUMOUR MARKERS
S. PSA
Urine R/M/E
S. ALPHA FETO PROTEIN
S. CEA
Selective Cases
S. C.A 19.9
USG of W/A
S. CA 125
MRI of Brain
S. Beta 2 Microglobulin
S. Protein Electrophoresis
S. LDH
CXR P/A View
ECG
ECHOCARDIOGRAM
OTHERS (If any)
37

ANNEXURE - D
(INVESTIGATIONS)
(2nd/3rd/4th/6th VISIT - 2/4/12/36 Weeks)
PARTICULARS OF THE PATIENT:
CODE NO:
NAME: 
AGE:
SEX:
M / F
REG. NO:
INVESTIGATIONS for Outcome Measures:
INVESTIGATIONS for Fitness & Maintainence:
CBC/ESR
BIOCHEMICAL:
Hb (gm/dl)
S.URIC ACID(umol/L)
ESR (mm in 1st hour)
S. Calcium
PLATELET COUNT (K/ul)
S. Phosphate
WBC 
Neutrophil (%)
S. ELECTROLYTES:
Lymphocyte (%)
SODIUM
Monocyte (%)
POTASSIUM
Eosinophil (%)
CHLORIDE
Basophil (%)
TCO2
ANION GAP
BIOCHEMICAL:
RBS
Urine R/M/E
HbA1C
S. Creatinine
e GFR (ml/min/1.73 m2)
BUN
Urine Microalbumin-Creatinine Ratio (UMCR)
38

ANNEXURE - D
(INVESTIGATIONS)
(5th / 7th VISIT - 24, 48 WEEK)
PARTICULARS OF THE PATIENT:
CODE NO:
NAME: 
AGE:
SEX:
M / F
REG. NO:
INVESTIGATIONS for Outcome Measures:
INVESTIGATIONS for Fitness & Maintainence:
DTPA Renogram
Total GFR
BIOCHEMICAL:
Right
S.URIC ACID(umol/L)
Left
S. Vit D3 (ng/ml)
S. Calcium
CBC/ESR
S. Phosphate
Hb (gm/dl)
ESR (mm in 1st hour)
S. ELECTROLYTES:
PLATELET COUNT (K/ul)
SODIUM
WBC 
POTASSIUM
Neutrophil (%)
CHLORIDE
Lymphocyte (%)
TCO2
Monocyte (%)
ANION GAP
Eosinophil (%)
Basophil (%)
BIOCHEMICAL:
RBS
HbA1C
S. Creatinine
e GFR (ml/min/1.73 m2)
BUN
Urine Microalbumin-Creatinine Ratio (UMCR)
Urinary Total Protein-Creatinine ratio(UPCR)
TUMOUR MARKERS
S. PSA (ng/ml)
S. ALPHA FETO PROTEIN (ng/ml)
S. CEA (ng/ml)
S. C.A 19.9 (U/ml)
S. CA 125
S. Beta 2 Microglobulin (mg/L)
S. LDH
39