Clinical Study of MSCs for Treatment of Poor Graft Function After Allogeneic Hematopoietic Stem Cell Transplantation
Phase: PHASE4
Conditions: MSC
Sponsor: ShiCang Yu

 
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Version: v3.0 
Date: December 12, 2023 
 
 
Mesenchymal Stem Cell Therapy for Hematopoietic 
Stem Cell Transplantation with Poor Graft 
Function: A Single-Center, Randomized, 
Controlled, Open Clinical Trial 
 
 
 
 
Principal Investigator: Gao Lei 
Research Institution: Second Affiliated Hospital of Army Medical University 
(formerly Third Military Medical University) 
Department: Hematology 
Participating Institution and Responsible Person: Chongqing Guolian Stem Cell 
Technology Co., Ltd., a subsidiary of Shanghai Saiao Biotechnology Co., Ltd., 
Shao Xiaoyan 
 
 
 
Confidentiality Statement: 
The information contained in this research proposal is provided solely for the 
review by the researchers, ethics committee, and relevant institutions of this 
project. Without the approval of the Principal Investigator (PI), it is strictly 
prohibited to disclose any information to third parties unrelated to this research. 
 
 
 
 

 
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Research Proposal Abstract 
Title 
Clinical Study on the Therapeutic Effect of Mesenchymal Stem Cells 
in the Treatment of Poor Graft Function after Allogeneic 
Hematopoietic Stem Cell Transplantation 
Applicant 
Second Affiliated Hospital of Army Medical University (formerly 
known as Third Military Medical University) 
Primary Research 
Institution 
Second Affiliated Hospital of Army Medical University (formerly 
known as Third Military Medical University) 
Research enter 
Second Affiliated Hospital of Army Medical University (formerly 
known as Third Military Medical University) 
Research 
Objective 
To explore the efficacy of umbilical cord-derived mesenchymal stem 
cells in the treatment of Poor Graft Function (PGF) after allogeneic 
hematopoietic stem cell transplantation. 
Research 
Hypothesis 
Umbilical cord-derived mesenchymal stem cells can treat impaired 
hematopoietic function after allogeneic hematopoietic stem cell 
transplantation. 
Research Design 
Single-center, randomized, controlled, open study. 
Inclusion Criteria 
(Main) 
1) Patients 
undergoing 
allogeneic 
hematopoietic 
stem 
cell 
transplantation. 
2) Both genders, aged ≥18 and ≤60 years. 
3) Karnofsky Performance Status (KPS) score >60, predicted 
survival >3 months. 
4) No severe systemic organ dysfunction. 
5) No contraindications for other hematopoietic stem cell 
transplantation. 
6) Voluntary participation with informed consent. 
 
Exclusion 
Criteria (Main) 
1) Severe heart, kidney, or liver dysfunction. 
2) Patients requiring treatment for other malignant tumors. 
3) Clinical symptoms of brain dysfunction or severe psychiatric 
illness that impairs understanding or adherence to the research 
protocol. 
4) Inability to complete the necessary treatment plan and follow-up 
observations. 
5) Patients with severe acute allergic reactions. 
6) Clinically uncontrolled active infection. 
7) Patients currently participating in other clinical trials. 
8) Other reasons deemed unsuitable for the clinical trial by the 
investigators. 
Simple size 
The ratio of cases between the experimental group and the control 
group is 1:1. Each group requires 31 cases, considering a possible 
dropout rate of 10%, resulting in a total of 34 cases per group. 
Intervention 
In addition to routine treatment for poor graft function, the 
experimental group will receive weekly infusion of umbilical cord-
derived mesenchymal stem cells at a dose of 1×10^6/kg for four 
consecutive weeks. 
Efficacy 
Indicators 
Primary endpoint: Recovery of blood parameters in patients with poor 
graft function after treatment. 
Secondary endpoints: Infection rate, graft-versus-host disease, 

 
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survival status (overall survival rate and disease-free survival rate 
within 1 year). 
Safety Indicators 
Incidence of other transplantation-related complications (infection, 
graft dysfunction, etc.) within 1 year after transplantation, acute 
adverse events, rejection reactions, and relapse rate within 1 year 
after transplantation. 
Expected 
Timeline 
September 2023 to December 2023: Design a single-center, 
randomized clinical research protocol and conduct its validation. 
January 2024 to December 2024: Initiate the clinical study, 
including ethics evaluation and international clinical research 
registration. Conduct a single-center, randomized clinical study on 
MSCs therapy for PGF, accumulate cases, and conduct mechanistic 
research. Prepare one research paper. 
January 2025 to October 2026: Continue the single-center, 
randomized clinical study on MSCs therapy for PGF, obtain 
midterm efficacy and safety data. Summarize the midterm data and 
prepare one research paper. 
November 2027 to September 2028: Complete the trial enrollment, 
continue follow-up, and summarize the clinical research data. Prepare 
one to two research papers. 

 
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1. Research Background 
Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is an effective 
treatment for benign and malignant hematological disorders. It mainly involves high-
dose chemotherapy to eliminate tumor cells, restore the recipient's hematopoiesis and 
immune function, and replace the recipient's hematopoietic stem cells (HSCs) with 
donor-derived HSCs to rebuild the donor's hematopoietic and immune function, 
thereby achieving the goal of curing the disease [1]. Successful engraftment of 
hematopoietic stem cells is characterized by the recovery of neutrophils, red blood 
cells, and platelets. Neutrophil engraftment is defined as a continuous presence of 
neutrophils exceeding 0.5×109/L for three consecutive days; platelet engraftment is 
defined as a continuous platelet count of at least 20×109/L for seven consecutive days 
without transfusion; and red blood cell engraftment is defined as a hemoglobin level 
of at least 70g/L without the need for blood transfusion [2-3]. Poor graft function 
(PGF) after transplantation is one of the main factors limiting the improvement of 
allo-HSCT efficacy. Therefore, it is of great clinical significance to explore the factors 
influencing successful engraftment of hematopoietic stem cells and new treatment 
strategies. 
 
Mesenchymal stromal cells (MSCs) were first identified by Friedenstein in the 
bone marrow stroma in 1968 as a non-hematopoietic adult multipotent stem cell [4]. 
It has been found that MSCs can maintain and increase the specific colony-forming 
units of CD34+ hematopoietic stem cells [5], participate in the regulation of 
hematopoietic cell growth, and also have unique immunomodulatory characteristics. 
MSCs express moderate levels of human leukocyte antigen (HLA) class I molecules, 
do not express HLA class II molecules, FAS ligand, or co-stimulatory molecules such 
as B7-1, B7-2, and CD40L. They can inhibit the proliferation response of T cells in 
mixed lymphocyte cultures (MLCs) or mitogen stimulation. Animal experiments 
have shown that infusion of HLA-matched or "third-party" MSCs expanded in vitro 
can prolong the survival of allogeneic skin grafts [6], confirming the 
immunosuppressive effects of MSCs in vivo. 
 
Risk factors for PGF after allo-HSCT include graft-versus-host disease (GVHD), 
the number of hematopoietic stem cells, anti-HLA antibodies (donor-specific 
antibodies) [7], previous alloimmune reactions, conditioning regimen intensity, 
cytomegalovirus (CMV) infection, graft source, low platelet count before 
conditioning, iron overload, etc. [8]. Currently, there is a lack of standardized 
treatment protocols for PGF after allo-HSCT, and treatment mainly focuses on 
addressing the underlying causes such as viral and infectious agents, hormones, etc. 
The intervention period is long, and the effective rate is less than 40%. Therefore, 
there is an urgent need to explore new and effective treatment methods. Multiple 
studies and preliminary research by our research team have indicated that 
mesenchymal stem cells (MSCs) can improve the bone marrow hematopoietic 
microenvironment, promote the proliferation and differentiation of hematopoietic 
stem cells, enhance hematopoietic function, and support hematopoiesis, as well as 

 
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directly or indirectly promote vascular regeneration in injured tissues and organs by 
secreting various cytokines. Existing research has confirmed a close association 
between the occurrence of PGF and endothelial damage in the bone marrow 
microenvironment, MSC damage, and imbalanced oxygen metabolism-induced HSC 
damage [9]. Therefore, exploring the treatment of PGF after transplantation from the 
perspective of improving the bone marrow hematopoietic microenvironment has 
certain feasibility. Currently, there is limited research on MSC treatment for PGF, and 
there are no reports on the use of systematic treatment protocols for PGF. Based on 
our previous research, we plan to adopt a safe and effective MSC treatment protocol 
for PGF and preliminarily elucidate its mechanism. 
 
With the support of the National Key Research and Development Program Stem 
Cell Research and Organ Repair Major Project Subproject (2022YFA1103300, 
2022YFA1103304), we have designed a single-center, prospective, randomized, 
controlled clinical study. We have also consulted renowned domestic experts in 
hematopoietic stem cell transplantation multiple times to develop the research 
protocol. For allo-HSCT patients, we will use 4th generation UC-MSCs, which have 
not undergone any genetic modification or gene editing. Based on conventional 
treatment for poor engraftment, umbilical cord blood-derived mesenchymal stem 
cells will be infused at a dose of 1×10^6/kg per week for four consecutive weeks. We 
aim to explore the efficacy of umbilicalcord blood-derived mesenchymal stem cells 
(UC-MSCs) in improving the engraftment of hematopoietic stem cells and the 
treatment of poor graft function (PGF) after allogeneic hematopoietic stem cell 
transplantation (allo-HSCT). 
 
The objective of this clinical study is to evaluate the safety and efficacy of UC-
MSCs in improving the engraftment of hematopoietic stem cells and treating PGF 
after allo-HSCT. The study will be conducted as a single-center, prospective, 
randomized, controlled trial. 
 
The study design involves two groups: the UC-MSC treatment group and the 
control group. Patients in the UC-MSC treatment group will receive UC-MSC 
infusions at a dose of 1×10^6/kg per week for four consecutive weeks in addition to 
conventional treatment for poor engraftment. The control group will receive only 
conventional treatment. 
 
The primary outcome measures of the study include the rate of neutrophil 
engraftment, platelet engraftment, and red blood cell engraftment. Secondary 
outcome measures include the incidence of graft failure, the time to neutrophil and 
platelet engraftment, the incidence of acute graft-versus-host disease (GVHD), 
overall survival, and other relevant clinical outcomes. 
 
The study will also assess the safety of UC-MSC treatment by monitoring adverse 
events, laboratory parameters, and immune-related parameters. 

 
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The research team plans to enroll a sufficient number of patients to achieve 
statistically significant results. Data analysis will be performed using appropriate 
statistical methods to compare the outcomes between the UC-MSC treatment group 
and the control group. 
 
The study protocol has been developed in consultation with experts in the field and 
has received support from the National Key Research and Development Program 
Stem Cell Research and Organ Repair Major Project Subproject. 
 
By investigating the use of UC-MSCs in the treatment of PGF after allo-HSCT, 
this study aims to contribute to the development of new treatment strategies for 
improving the engraftment of hematopoietic stem cells and enhancing the outcomes 
of allo-HSCT. The results of this study may have implications for the clinical 
management of patients undergoing allo-HSCT and provide insights into the potential 
therapeutic applications of UC-MSCs in hematopoietic stem cell transplantation. 
Reference： 
[1] Xu L , Chen H , Chen J , et al. The consensus on indications, conditioning regimen, 
and donor selection of allogeneic hematopoietic cell transplantation for hematological 
diseases in China—recommendations from the Chinese Society of Hematology[J]. 
Journal of Hematology & Oncology, 2018, 11(1):33. 
[2] KONG Y, SONG Y, TANG F F, et al. N-acetyl-L-cysteine improves mesenchymal 
stem cell function in prolonged isolated thrombocytopenia post-allotransplant [J]. Br J 
Haematol, 2018,180(6): 863-78. 
[3]First LR,Smith BR,Lipton J,et al.Isolated thrombocytopenia after allogeneic bone 
marrow transplantation:existence of transient and chronic thrombocytopenic 
sumdromes[J],Blood,1985,65:368-374. 
[4]Friedenstein AJ, Petrakova KV, Kurolesova AI, et al. Hetero typic transplants of  
bone marrow: analysis of precursor cells for osteogenic and hematopoietic tissues. 
Transplantation, 1968, 6 (2): 230-247. 
[5]Cheng L , Qasba P, Vanguri P, et al. Human mesenchymal stem cells support 
megacaryocyte and prop latelet formation from CD34+ hematopoietic progenitor cells. 
J Cell Physiol, 2000, 184 (1) : 58-59. 
[6]Krampera M , Glennie S, Dyson J, et al. Bonemarrow mesenchymal stem cells 
inhibit the response of naive and memory antigenspecific T cells to their cognate 
peptide. Blood, 2003, 101 (9): 3722-3729. 
[7] MAHAT U, ROTZ S J, HANNA R. Use of thrombopoietin receptoragonists in 
prolonged thrombocytopenia after hematopoietic stem cell transplantation [J]. Biol 
Blood Marrow Transplant,2020, 26(3): e65-73. 
[8] Michalicka M，Boisjoli G,Jahan S，et al．Human bone marrow mesenchymal 
stromal cell-derived osteoblasts promote the expansion of hematopoietic progenitors 

 
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through betacatenin and notch signaling pathways［J］．Stem Cells Dev,2017,26( 24) : 
1735-1748. 
[9] Wojciech Jurczak , Krzysztof Chojnowski ,et al，Phase 3 randomised study of 
avatrombopag, a novel thrombopoietin receptor agonist for the treatment of chronic 
immune thrombocytopenia[J],Br J Haematol.2018;183(3):479-490. 
 
 
2.Clinical Trial Objective 
To explore the efficacy, safety, and related mechanisms of umbilical cord-derived 
mesenchymal stem cell therapy in the treatment of hematopoietic graft failure and 
engraftment dysfunction after allogeneic hematopoietic stem cell transplantation. 
 
3.Clinical Trial Design 
This is a single-center, randomized, controlled, open-label clinical trial designed to 
primarily investigate the efficacy and safety of umbilical cord-derived mesenchymal 
stem cell therapy in the treatment of engraftment dysfunction after allogeneic 
hematopoietic stem cell transplantation. 
 
4.Study Population 
4.1 Number of Cases 
A total of 68 patients will be enrolled, with 34 patients in the experimental group and 
34 patients in the control group. 
4.2 Inclusion Criteria 
The following criteria must be met for patient inclusion: 
 
Patients undergoing allogeneic hematopoietic stem cell transplantation. 
Both genders, aged ≥18 and ≤60. 
Karnofsky Performance Status (KPS) score >60, estimated survival period >3 months. 
No severe systemic impairment of vital organ function. 
No contraindications to other hematopoietic stem cell transplantation. 
Voluntary participation and informed consent. 
4.3 Exclusion Criteria 
Patients meeting any of the following criteria are not eligible for inclusion: 
Severe heart, kidney, or liver dysfunction. 
Patients requiring treatment for other malignancies. 
Clinical symptoms of brain dysfunction or severe psychiatric disorders that affect 
comprehension or compliance with the study protocol. 
Patients unable to complete the required treatment plans and follow-up observations. 
Patients with severe acute allergic reactions. 

 
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Clinically uncontrolled active infections. 
Patients currently participating in other clinical trials. 
Participants deemed unsuitable for the clinical trial by the researchers. 
4.4 Criteria for Removal or Dropout: 
Patients experiencing disease progression unrelated to the experimental factors or death 
during the treatment and follow-up period will be unable to continue the observation. 
4.5 Termination Criteria: 
Occurrence of severe adverse reactions or intolerability. 
Participant's voluntary withdrawal. 
Disease progression or relapse during the trial period. 
Participants deemed unsuitable for further treatment by the researchers. 
5.Treatment Protocol 
5.1 Treatment for Post-Transplantation Poor Graft Function (PGF) after Allogeneic 
Hematopoietic Stem Cell Transplantation 
Experimental Group: UC-MSCs (umbilical cord-derived mesenchymal stem cells) for 
infusion will be provided by Chongqing Guolian Stem Cell Technology Co., Ltd., a 
subsidiary of Shanghai Saiao Biotechnology Co., Ltd. The umbilical cord tissue used 
as the source is obtained legally and meets ethical requirements. The UC-MSCs are not 
genetically modified or edited and undergo strict standard procedures and quality 
control measures during isolation, culture, testing, and transportation, ensuring their 
safety. The technologies "Clinical-Grade Umbilical Cord Mesenchymal Stem Cells and 
Method for Their Isolation and Preparation" and "Method for Isolation and Culture of 
Umbilical Cord Tissue-Derived Mesenchymal Stem Cells and Products Thereof" 
developed by Chongqing Guolian Stem Cell Technology Co., Ltd., a subsidiary of 
Shanghai Saiao Biotechnology Co., Ltd., have been granted patents by the National 
Intellectual Property Administration of China. The company has the qualification for 
UC-MSCs preparation, and the UC-MSCs prepared by them have been certified as 
qualified by a third-party testing institution. Therefore, UC-MSCs provided by 
Chongqing Guolian Stem Cell Technology Co., Ltd. will be used in this study. The 
preparation fee for UC-MSCs will be provided by the research funds of the National 
Key Research and Development Program "Stem Cell Research and Organ 
Regeneration," and the staff of the UC-MSCs preparation company will not participate 
in this study. In addition to routine PGF treatment, patients in the experimental group 
will receive umbilical cord blood-derived mesenchymal stem cells at a dose of 
1×10^6/kg once a week for four consecutive weeks. 
Control Group: Standard PGF treatment, including red blood cell and platelet 
transfusions, subcutaneous injection of granulocyte colony-stimulating factor (G-CSF) 
at a dose of 5-10ug/kg/day, subcutaneous injection of recombinant human 
thrombopoietin injection (TPO) at a dose of 300U/kg/day, and TPO receptor agonist 

 
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(TPO-RA) (starting dose of 1 tablet once daily, PLT<50×10^9/L for at least 2 weeks, 
increase by one dose level, PLT 200-400×10^9/L, decrease by one dose level, 
PLT>400×10^9/L, discontinue TPO-RA). 
5.2 Observational IndicatorsI apologize, but the information you provided seems to be 
cut off.  
 
6.Research Procedure 
6.1 Recruitment and Baseline Evaluation  
Subjects who meet the conditions of Section 4.1 of this research protocol will 
undergo a series of necessary functional examinations as essential research 
indicators for clinical studies. The main examination items include but are not 
limited to the following: 
a. Physical examination (weight, height, surface area) 
b. Vital signs examination: temperature, pulse, cardiac function 
c. Complete blood count: hemoglobin, white blood cells, platelets, etc. 
d. Liver and kidney function tests 
e. Bone marrow examination, bone marrow biopsy 
f. Minimal Residual Disease (MRD) and related gene mutation detection 
g. Lymphocyte subset analysis 
h. Imaging examinations: ultrasound, CT/MRI/DSA, etc. 
6.2 Subject Document Collection 
The documents required for subject recruitment include: 
 
Completion of screening materials; 
Signing the informed consent form; 
6.3Implementation of Allogeneic Hematopoietic Stem Cell 
Transplantation 
6.3.1 Preparative Regimen: 
The preparative regimen for Haplo-HSCT patients is: Me-CCNU+Ara-
C+Bu+Cy+ATG 
Lomustine (Me-CCNU): 250mg/m2×1d 
Cytarabine (Ara-C): 4g/m2×2d 
Busulfan (Bu): 0.8mg/kg q6h, ×2d 
Cyclophosphamide (CTX): 1.8g/m2×2d 
Anti-thymocyte globulin (ATG): 2.5mg/kg×4d 
 
The preparative regimen for SMD-HSCT patients is: Bu+Cy+ATG 
Busulfan (Bu): 0.8mg/kg q6h×4d 

 
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Cyclophosphamide (CTX): 60mg/kg×2d 
 
The preparative regimen for UMD-HSCT patients is: BU+CY+ATG 
Busulfan (Bu): 0.8mg/kg q6h×4d 
Cyclophosphamide (CTX): 60mg/kg×2d 
Anti-thymocyte globulin (ATG): 2.5mg/kg×4d 
6.3.2 Mobilization, Collection, and Infusion of Hematopoietic Stem 
Cells 
Recombinant human granulocyte colony-stimulating factor (rhG-CSF) is 
administered subcutaneously at a dose of 10ug/kg/day for 4 days. On day 5, 
peripheral blood stem cells (PBSC) are collected using the German Fresenius 
blood cell separator. On day 6, bone marrow aspiration is performed under 
epidural anesthesia. The donor lies prone on the operating table, and bone marrow 
is collected by puncturing at different depths and multiple sites on both iliac crests. 
If more than 600ml of bone marrow fluid is expected to be collected, the donor 
needs to be adjusted to a supine position, and bone marrow is collected by 
puncturing at the anterior superior iliac spine on both sides. If there is ABO blood 
type mismatch between the donor and recipient, hydroxyethyl starch 
sedimentation is used to remove red blood cells. If there is minor ABO blood type 
mismatch, the bone marrow fluid needs to be centrifuged at 4℃ and then plasma 
is removed using a plasma extractor. 
6.4 Treatment of Post-Transplantation Graft Failure (PGF) 
Experimental group: UC-MSCs (Umbilical Cord Mesenchymal Stem Cells) 
administered without any genetic modification or editing. In addition to routine 
treatment for PGF, umbilical cord mesenchymal stem cells are infused at a dose of 
1×10^6/kg once a week for 4 consecutive weeks. 
Control group: Conventional treatment for PGF, including red blood cell and 
platelet transfusion, subcutaneous injection of granulocyte colony-stimulating 
factor (G-CSF) at a dose of 5-10ug/kg/day, subcutaneous injection of recombinant 
human thrombopoietin injection (TPO) at a dose of 300U/kg/day, and TPO 
receptor agonist (TPO-RA) administration (starting dose of 1 tablet, once daily, 
for at least 2 weeks if PLT<50×10^9/L, increasing one dose level if PLT reaches 
200-400×10^9/L, decreasing one dose level if PLT>400×10^9/L, discontinuing 
TPO-RA). 
6.5 Observations of Indicators 
Before hematopoietic reconstitution: Complete blood count (twice a week), liver 
and kidney function tests, coagulation function tests (weekly),cardiac function 
tests (as needed), and imaging examinations (as needed). 

 
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After hematopoietic reconstitution: Complete blood count (twice a week for the 
first month, then once a week for the second month, and once every two weeks for 
the third month), lymphocyte subset analysis (once a month for the first year), 
minimal residual disease (MRD) and related gene mutation detection (as needed), 
and imaging examinations (as needed). 
Post-transplantation graft failure (PGF) observation: Complete blood count (daily), 
blood culture (as needed), liver and kidney function tests (as needed), coagulation 
function tests (as needed), and imaging examinations (as needed). 
Long-term follow-up: Complete blood count (once every three months for the first 
year, then once every six months for the second year, and once a year thereafter), 
liver and kidney function tests (once every three months for the first year, then 
once every six months for the second year, and once a year thereafter), coagulation 
function tests (once every three months for the first year, then once every six 
months for the second year, and once a year thereafter), lymphocyte subset 
analysis (once a year), minimal residual disease (MRD) and related gene mutation 
detection (as needed), and imaging examinations (as needed). 
 
 
6.6 Treatment Monitoring Plan 
Pre-hematopoietic Reconstruction Monitoring Indicators: 
(1) Changes in body temperature, pulse, and blood pressure (daily) 
(2) Complete blood count (twice a week) 
(3) Liver, kidney, and coagulation function tests (weekly) 
(4) CMV, EB, and other viral indicators (1-2 times a week) 
From hematopoietic reconstruction to 1 year post-transplant: 
(1) Complete blood count, liver and kidney function tests, CMV and EB, and other 
viral indicators (at least monthly) 
(2) Bone marrow examination, bone marrow biopsy (monthly) 
(3) MRD (minimal residual disease), detection of relevant gene mutations 
(monthly) 
(4) Lymphocyte subset analysis (1 month, 2 months, 3 months, 4.5 months, 6 
months, 9 months, 12 months after transplantation) 
 
6.7 Assessment Criteria for Treatment Efficacy 
White blood cells: Neutrophil engraftment is defined as neutrophil count exceeding 
0.5×10^9/L for three consecutive days. 
Red blood cells: Hemoglobin level should be no lower than 70g/L, and the patient 
should no longer require blood transfusions. 
Platelets: 

 
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Complete response (CR) is defined as a platelet count ≥50×10^9/L for seven 
consecutive days without platelet transfusion. 
Partial response (PR) is defined as a platelet count between 20×10^9/L and 
50×10^9/L for seven consecutive days without platelet transfusion. 
No response (NR) is defined as a platelet count <20×10^9/L or failure to discontinue 
platelet transfusion after eight weeks of treatment at the maximum tolerated dose. 
 
6.8 Criteria for Assessing the Efficacy of Acute Leukemia Treatment 
The evaluation of acute leukemia treatment is based on the 2017 edition of the 
"Chinese Clinical Diagnosis and Treatment Guidelines for Adult Acute Myeloid 
Leukemia (Non-Acute Promyelocytic Leukemia)" and the 2016 edition of the 
"Chinese Diagnosis and Treatment Guidelines for Adult Acute Lymphoblastic 
Leukemia." The assessment results may include the following possibilities: complete 
remission (morphological complete remission, molecular complete remission, 
morphological complete remission with incomplete blood cell recovery), relapse 
(molecular/genetic relapse, hematological relapse). 
 
 
 
7. Safety and Adverse Events 
7.1 Definition 
Unexpected problems that may occur in patients or others: Any event, 
experience, or difficulty that meets the following conditions, including: unexpected 
nature, severity, or frequency (such as the absence of mentioned approvals from the 
Institutional Review Board (IRB) in study-related documents, protocols, informed 
consent forms, investigator manuals, etc.); related or potentially related to 
participation in the study (potential relevance means that there may be unforeseen 
events or results affecting the trial protocol in this study); indicating that the 
researcher puts patients or others at significant risk (including physical, psychological, 
economic, or social harm). 
 
Adverse Event (AE): An adverse event is a symptom, sign, disease, or other 
development that reaches a severe level during the study process. Complications and 
injuries are considered adverse events. Abnormal diagnostic results that meet the 
following criteria are also considered adverse events: withdrawal from the study, 
association with many serious adverse events, association with clinical signs and 
symptoms, leading to additional treatment or further diagnostic tests, considered 
clinically significant by the study subjects. 
 
Serious Adverse Event: Adverse events are categorized as serious or non-serious. 
Serious adverse events refer to the following adverse events: fatal, life-threatening, 

 
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requiring or prolonging hospitalization, causing persistent or severe disability, and 
other significant medical events. 
 
Significant medical events refer to events that may not immediately threaten life 
but have significant clinical significance. It may jeopardize the patient or may require 
intervention to prevent the occurrence of other serious events mentioned above. For 
example, drug overdose or abuse, sudden onset of a disease that does not require 
hospitalization of the patient, intensification of bronchospasm treatment in the 
emergency department is usually considered serious. All other adverse events that do 
not meet the criteria for a serious adverse event are considered non-serious adverse 
events. 
 
Dose-Limiting Toxicity (DLT): Dose-limiting toxicity is defined as Grade 3 or 
Grade 4 toxicity. Additionally, if the toxicity is autoimmune or natural allergy and 
occurs after repeated administration, Grade 2 toxicity is also considered DLT. 
 
Adverse Event Reporting Period: The period during which adverse events 
occurring during the study need to be reported generally refers to the time from the 
start of the study to the end of the follow-up of study treatment. For this study, the 
follow-up of study treatment occurs within 6 months after the implementation of the 
last treatment. Adverse events occurring during chemotherapy and outcomes directly 
caused by antitumor chemotherapy are not within the adverse event reporting period. 
 
Pre-Existing Condition: Pre-existing conditions refer to conditions that already 
exist at the start of the study. Pre-existing conditions need to be documented because 
frequent, severe, or worsening of pre-existing diseases may occur during the study 
process. 
 
General Medical Findings: During the screening process, any clinically 
significant abnormalities are recorded as pre-existing conditions. After the study ends, 
any new clinically significant findings/abnormalities that meet the definition of 
adverse events must be recorded as adverse events. 
 
Post-Study Adverse Events: All unresolved adverse events should be followed 
up by the investigator until they are resolved unless the patient is lost to follow-up or 
there is another explanation for the adverse event. At the last scheduled visit, the 
investigator needs to instruct each patient to report any events they or their personal 
physician believe may be related to their participation in the study. The investigator 
should note the occurrence of death events in the study or adverse events that occur 

 
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after some patients discontinue the study or terminate their participation in the study. 
The investigator should also note the possibility of cancer development in patients 
participating in the study or the presence of congenital abnormalities in offspring 
born to study participants. 
 
Laboratory Abnormalities: If any of the following conditions occur in clinical 
laboratory abnormalities, they will be recorded as adverse events: abnormalities 
observed in the laboratory that have been proven to be abnormal through repeated 
testing and cannot be refuted in terms of their abnormality, abnormalities indicating 
the presence of diseases and/or organ toxicity. Abnormal levels need to be actively 
managed, such as dose adjustments, discontinuation of medication, more frequent 
follow-up assessments, further diagnostic investigations, etc. Laboratory 
abnormalities attributed to potential diseases and chemotherapy regimens are not 
considered adverse events (such as abnormal blood indices and mucositis). 
 
Hospitalization, Prolonged Hospitalization, or Surgery: Any adverse event that 
leads to hospitalization or prolonged hospitalization should be recorded as a serious 
adverse event unless there are specific explanations in this protocol. Any condition 
responsible for surgery that meets the criteria for an adverse event needs to be 
recorded as an adverse event. As mentioned above, events related to antitumor 
chemotherapy are excluded from adverse events. Whether it is a condition, 
hospitalization, prolonged hospitalization, or surgery, it needs to be recorded as an 
adverse eventThe information provided above outlines the definitions and criteria for 
safety and adverse events in a clinical study. It includes the definition of adverse 
events, serious adverse events, dose-limiting toxicity, adverse event reporting period, 
pre-existing conditions, general medical findings, post-study adverse events, 
laboratory abnormalities, and hospitalization or surgery related to adverse events. 
 
In summary, safety and adverse events are closely monitored during a clinical 
study to ensure the well-being of participants. Any unexpected problems, symptoms, 
diseases, or developments that occur during the study are recorded and evaluated. 
Adverse events are categorized as serious or non-serious, with serious adverse events 
encompassing severe or life-threatening events that may require hospitalization or 
cause significant medical consequences. 
 
The reporting period for adverse events typically extends from the start of the 
study to the end of the follow-up of study treatment. Pre-existing conditions and any 
new clinically significant findings or abnormalities discovered during the study are 
documented as adverse events. Post-study adverse events, unresolved adverse events, 

 
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or events occurring after a participant's discontinuation of the study are also followed 
up and recorded. 
 
Laboratory abnormalities indicating diseases, organ toxicity, or abnormalities 
that require active management are considered adverse events. Hospitalization, 
prolonged hospitalization, or surgery resulting from adverse events are also 
documented. 
 
It's important to note that the information provided is a general overview, and 
specific protocols and procedures may vary depending on the clinical study and its 
objectives. 
 
7.2 Adverse Event Recording 
During communication with each patient, the investigator can collect information on 
adverse events through specific inquiries or, in certain specific situations, through 
testing. All information regarding adverse events needs to be immediately entered 
into the database and recorded in the Adverse Event module of the Case Report Form 
(CRF). All evident relevant signs, symptoms, and abnormal diagnostic results need 
to be entered into the database and grouped under the same diagnosis. 
 
All adverse events occurring during the study period need to be recorded. All adverse 
events need to be tracked until resolution, stabilization, or confirmation that the study 
treatment or participation is not an issue. Serious adverse events that persist beyond 
the end of the study must be tracked until final resolution is determined. Any serious 
adverse events occurring after the study that are considered possibly related to the 
study treatment or participation need to be recorded and promptly reported. 
 
7.3 Toxicity Management, Termination Rules, and Study Termination 
7.3.1 Criteria for Study Termination or SuspensionThe study will be terminated 
if the following conditions occur: 
 
If a subject experiences a Grade 3 or higher toxicity reaction following cell infusion 
therapy, identified as originating from the treatment protocol of this clinical trial, the 
clinical protocol will be modified to reduce the dosage of the treatment drug based 
on the nature of the toxicity reaction. 
If the investigator or any independent review committee or regulatory authority 
determines that this trial would pose a safety risk to the subjects, the clinical trial will 
be terminated. 
If the investigator decides to terminate this clinical trial, the trial will be terminated. 

 
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If any subject experiences the following conditions within 2 weeks after receiving 
cell infusion therapy, the study will be required to undergo reevaluation, and the 
clinical trial at that stage will be suspended: 
 
Respiratory failure requiring mechanical ventilation. 
Grade 4 (life-threatening) toxicity reaction resulting from the treatment protocol. 
Death. 
7.3.2Manegement of Toxic Effects 
Infusion reactions: No literature reports of infusion reactions with MSCs have 
been observed. If they occur, anti-allergic and symptomatic treatments such as 
dexamethasone will be administered。 
 
7.3.3 Criteria for Terminating Subject Participation in the Clinical Trial 
If, after recruitment into the clinical trial, a subject cannot undergo allogeneic 
hematopoietic stem cell transplantation for various reasons, the subject will be 
terminated from the trial. This decision will be made by the principal investigator and 
may include factors such as changes in the underlying disease, severe infection, or 
significant organ dysfunction. Additionally, if relapse occurs during the MSC 
infusion period (within 12 weeks after transplantation), MSC preventive treatment 
will be terminated. 
8.Sample Size Estimation 
From the perspective of sample composition, the study is divided into the 
experimental group (MSC treatment group) and the control group. The estimation of 
sample size for one-sided testing using the two-sample rate comparison difference 
testing formula is calculated as c = n1/n2, with a statistically significant difference 
set at 0.05 and a detection power (P) value set at 0.80. The incidence rates in the 
experimental and control groups are 76% and 38%, respectively (based on previous 
historical control study results). Plugging these values into the formula yields a 
required sample size of 31 cases per group. Considering a possible dropout rate of 
10%, each group will require 34 cases. The ratio of cases between the experimental 
and control groups is 1:1. 
 
 
9.Data Processing and Record Keeping 
9.1 Principle of Confidentiality 
All information from the clinical trial is highly confidential. Except for inspections 
and duplication of records by regulatory authorities and sponsors or their 
representatives, no unauthorized personnel should be provided access to the 
information without written authorization from the sponsor. Unless authorized in 

 
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writing by the sponsor, all investigational drugs, subjects' biosamples, or other 
collected materials must be used according to the trial protocol. 
In the case report form, the subjects' identities will be recorded using the uppercase 
initials of their names and unique subject codes. 
9.2 Document Storage 
To ensure evaluation and supervision by the National Medical Products 
Administration and the sponsor, the investigator should store all data from this 
clinical trial in accordance with relevant national regulations. This includes 
confirmation of all subjects (to effectively cross-check different records, such as 
CRFs and original hospital records and laboratory examination reports), all signed 
informed consent forms, all CRFs, and detailed records of drug distribution. The 
storage period is 5 years. 
The sponsor should retain the trial protocol, ethics committee approvals, monitoring 
records, contracts, and records of serious adverse events for the required period as 
specified by the national drug regulatory authority. 
Under no circumstances should the investigator provide any form of trial-related 
information to third parties. 
9.3 Protocol Amendments 
Once the trial protocol is determined and approved by the ethics committee, the 
research design should generally not be changed. However, modifications may be 
considered in the following two scenarios: 
 
During the course of the trial, if it is found that it isnecessary to modify the protocol 
to ensure the safety and well-being of the subjects or to improve the scientific validity 
of the study, the investigator should submit a formal request for protocol amendment 
to the ethics committee for review and approval. 
In exceptional circumstances, urgent modifications to the protocol may be required 
to eliminate immediate hazards to the subjects. In such cases, the investigator should 
immediately inform the ethics committee and obtain approval for the modification. 
The sponsor should also be notified promptly. 
Protocol amendments should be documented and kept on file. All subsequent study 
procedures should adhere to the amended protocol, and the reasons for the 
amendments should be clearly stated in the final study report. 
10. Statistics Analysis 
Statistical analysis should be conducted by a qualified statistician using appropriate 
statistical methods. The analysis plan should be pre-specified in the protocol or a 
separate statistical analysis plan. 
Descriptive statistics should be used to summarize baseline characteristics of the 
study population, including demographics, disease characteristics, and other relevant 

 
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variables. 
 
For the primary outcome and secondary outcomes, appropriate statistical tests should 
be performed to compare the treatment groups. The choice of statistical tests should 
be justified based on the nature of the outcome variables and the study design. 
Confidence intervals and p-values should be reported for the estimated treatment 
effects. 
 
For safety outcomes, adverse events should be summarized using appropriate 
descriptive statistics, including frequencies and percentages. Serious adverse events 
should be reported separately. 
 
All statistical analyses should be conducted using appropriate statistical software, and 
the results should be reported in a clear and transparent manner. 
 
11. Ethical Considerations 
The study should be conducted in accordance with ethical principles outlined in 
the Declaration of Helsinki and applicable national and international regulations. The 
protocol should be reviewed and approved by an independent ethics committee or 
institutional review board before the study begins. 
Informed consent should be obtained from all study participants or their legally 
authorized representatives. The informed consent process should provide clear and 
understandable information about the study, its risks and potential benefits, and the 
rights of the participants. 
 
Privacy and confidentiality of the participants should be protected throughout 
the study. Personal information should be handled in accordance with applicable data 
protection laws. 
 
The welfare and rights of the study participants should be prioritized. Any 
unexpected or serious adverse events should be promptly reported to the ethics 
committee, and appropriate actions should be taken to ensure the safety and well-
being of the participants.