A Randomized Controlled Study of Mesenchymal Stem Cells in the Treatment of Chronic Kidney Diseases
Phase: PHASE1
Conditions: Kidney Disease, Chronic, Diabetic Kidney Disease (DKD), Hypertensive Nephropathy
Sponsor: The First Affiliated Hospital of Air Force Medicial University

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Source and Number of the Project：
Randomized Controlled Study of Mesenchymal
Stem Cell Therapy for Chronic Kidney Disease
Principal Investigator: He Lijie
Department in Charge: Nephrology Department
Contact Number: 029-89661611
Implementing Unit: The First Affiliated Hospital of Air Force
Medical University
Study Duration: 2024-01-01 – 2027-08-31
Version Number: V3.0
Version Date: 2024-09-19

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PROTOCOL SYNOPSIS
Study Title
Randomized
Controlled
Study
of
Intravenous
Mesenchymal Stem Cell Injection for Chronic Kidney
Disease
Objective
To investigate the therapeutic effect of allogeneic umbilical cord
tissue-derived mesenchymal stem cell intravenous injection
therapy on chronic kidney disease.
Design
This trial is a prospective, randomized, controlled clinical trial.
Eligible subjects will be randomized in a 1:1 ratio to either the
experimental group or the control group. The experimental
group will receive a single intravenous injection of allogeneic
umbilical cord tissue-derived MSCs（mesenchymal stem cells）
at a dose of 1.0*106/kg. Baseline data will be collected before
treatment. Follow-up assessments will be conducted at 1, 3, 6,
and 12 months post-treatment to evaluate the efficacy for
CKD(chronic kidney disease) patients (Stage 3 and 4).
Sample Size
32 subjects
Subject
Selection
Inclusion Criteria:
(1) Agreement to participate in the trial and provision of
signed written informed consent;
(2) Pathological diagnosis of diabetic nephropathy or
hypertensive renal damage;
(3) 15 ≤eGFR(estimated glomerular filtration rate) ＜60
mL/min/1.73m²,
UACR(urinary
albumin-to-creatinine
ratio) > 300 mg/g;
(4) Age ≥18 years.
Exclusion Criteria:
(1) Extremely severe anemia (hemoglobin < 30 g/L);

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(2) Received blood product transfusion therapy within 1
month;
(3) Autosomal dominant or recessive polycystic kidney
disease (ADPKD);
(4) History of kidney transplant or other solid organ
transplant;
(5) Active systemic or localized infection (e.g., pneumonia,
osteomyelitis);
(6) Allergy to stem cells themselves or stem cell-related
culture medium;
(7) History of allergic reaction to cell products (e.g., blood
transfusion, platelets);
(8) History of coagulation disorders (thromboembolism,
pulmonary embolism, deep vein thrombosis);
(9) History of malignancy or current malignant disease;
(10) Elevated tumor markers (AFP, CEA, CA199, CA125,
etc.);
(11) Pregnant women or women with plans for pregnancy
within 3 months after MSC （mesenchymal stem cell ）
therapy;
(12) Participation in drug-related clinical trials within the
past 2 months;
(13) Any form of drug abuse, mental illness, or other
conditions considered by the investigator as potentially
affecting the trial's validity or the subject's health.
Intervention
Eligible subjects will be randomized 1:1 to the experimental
group or the control group. The experimental group will receive
a single intravenous injection of allogeneic umbilical cord
tissue-derived MSCs at a dose of 1.0*106/kg. The control group

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will
receive
a
placebo.
Baseline
data
will
be
collected
pre-treatment.
Follow-ups
at
1,
3,
6,
and
12
months
post-treatment will guide subjects to return to the hospital for
relevant laboratory tests. The primary endpoints are estimated
glomerular
filtration
rate
(eGFR)
and
urinary
albumin-to-creatinine
ratio
(UACR).
Secondary
endpoints
include other renal function parameters, other organ system
function parameters, and MSC-related adverse events. The
MSCs for treatment will be provided by Shaanxi Zhonggang
Wanhai Institute of Life Sciences Co., Ltd. or Shaanxi Yisaier
Biotechnology Co., Ltd.
Endpoints
Efficacy Endpoints: Changes in eGFR, 24-hour urine protein,
and UACR at 1, 3, 6, and 12 months will serve as primary
endpoints to assess renal function improvement. Changes in
other renal function parameters (e.g., quantitative urine protein,
urine red blood cells, serum creatinine, cystatin C),and changes
in
other
organ
system
function
parameters
(e.g.,
ECG/echocardiography, complete blood count, liver function,
NT-proBNP, parathyroid hormone, 25-dihydroxyvitamin D,
lymphocyte subsets) at 1, 3, 6, and 12 months will serve as
secondary endpoints.
Safety Endpoints: Incidence of adverse events.
Statistical
Methods
An EDC database will be established. All data and results will be
entered into the database. Statistical analysis will be performed
using SPSS software, including t-tests, Wilcoxon rank-sum tests,
chi-square
tests,
Spearman
correlation
analysis,and
Cox
regression analysis, etc.
Study Period
January 2024 – August 2027

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1. Background
Chronic Kidney Disease (CKD) refers to chronic structural and
functional abnormalities of the kidneys caused by various etiologies,
encompassing a group of diseases with different causes, pathologies, and
varying severities[1]. CKD is defined as the presence of kidney damage
markers or a glomerular filtration rate (GFR) below 60mL/(min·1.73m²)
for more than 3 months. Besides GFR, the severity of proteinuria is also
an important factor influencing the progression rate and outcomes of
CKD[2]. The top three etiologies of CKD in Chinese patients are primary
glomerulonephritis,
diabetic
nephropathy,
and
hypertensive
nephrosclerosis. CKD is a progressive, incurable disease with high
morbidity and mortality, posing a serious public health problem
worldwide.
Its
global
average
prevalence
is
9.1%,
affecting
approximately 697.5 million patients, with nearly 132 million in China[3].
Current treatment strategies include therapies to delay the progression of
chronic renal insufficiency and treatments for various complications,
namely general treatment, treatment of the primary disease, and measures
to slow the progression of chronic renal failure. When the disease
progresses to end-stage renal disease (ESRD), patients can only undergo
renal replacement therapy, including hemodialysis, peritoneal dialysis,
and kidney transplantation[4].

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Mesenchymal Stem Cells (MSCs) are multipotent adult stem cells
with self-renewal capacity, derived from the mesoderm. They have
become a focus of advanced biomedical technology worldwide and have
demonstrated safety and efficacy in clinical trials for various diseases[5],
even being included in expert consensus for liver disease treatment[6].
MSCs have the potential to promote regeneration of damaged renal tissue,
achieving renal protection and even reversal of injury, offering a new
possibility and supplement to existing therapies.
Currently, clinical trials of MSCs for kidney diseases have mainly
focused on lupus nephritis[7,
8], diabetic nephropathy[9-11], and acute
kidney injury[12]. Their safety has been consistently verified, but due to
generally small sample sizes and a lack of long-term follow-up, their
efficacy remains controversial[13]. Therefore, our research team plans to
conduct this randomized controlled clinical trial. Through long-term
follow-up and observation of changes in multiple renal and other organ
function parameters, we aim to further validate the therapeutic effect of
MSCs on patients with severe chronic kidney disease (CKD stages 3 and
4), providing new ideas and methods for the clinical treatment and
prognosis improvement of CKD patients.
2. Study Objectives
To evaluate the effect of intravenous injection of umbilical cord
tissue-derived mesenchymal stem cells on the improvement of renal
function in patients with chronic kidney disease (CKD stages 3 and 4),
using the change in estimated glomerular filtration rate (eGFR) as the
primary outcome measure, and changes in other renal function laboratory

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parameters, changes in other organ system function laboratory parameters,
and the incidence of adverse events as secondary outcomes. This trial
aims to further verify and evaluate the therapeutic effect of MSCs on
patients with severe chronic kidney disease (CKD stages 3 and 4),
providing new ideas and strategies for expanding clinical treatment
options, delaying progression, and improving the prognosis of CKD
patients.
3. Study Design
This study is a prospective, randomized controlled clinical trial
initiated by the team of Associate Professor He Lijie from the Nephrology
Department of Xijing Hospital. Eligible subjects will be randomized in a
1:1 ratio to the experimental group or the control group. The experimental
group will receive a single intravenous injection of allogeneic umbilical
cord tissue-derived MSCs at a dose of 1.0*106/kg. The control group will
receive a placebo. Baseline data will be collected before treatment.
Follow-up assessments will be conducted at 1, 3, 6, and 12 months
post-treatment, guiding subjects to return to the hospital for relevant
laboratory tests. The primary endpoints are the estimated glomerular
filtration rate (eGFR) and urinary albumin-to-creatinine ratio (UACR) at
1, 3, 6, and 12 months after treatment. Secondary endpoints include other
renal function parameters, other organ system function parameters, and
MSC-related adverse events. The MSCs for treatment are prepared by
Shaanxi Zhonggang Wanhai Institute of Life Sciences Co., Ltd. or
Shaanxi Yisaier Biotechnology Co., Ltd.
4. Subject Selection
4.1 Inclusion Criteria
(1) Agreement to participate in the trial and provision of signed,

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written informed consent;
(2) Pathological diagnosis of diabetic nephropathy or hypertensive renal
damage;
(3) 15≤eGFR＜60min·1.73m²，UACR＞300mg/g；
(4) Age ≥18 years.
4.2 Exclusion Criteria
(1) Extremely severe anemia (hemoglobin < 30 g/L);
(2) Received blood product transfusion therapy within 1 month;
(3) Autosomal dominant or recessive polycystic kidney disease (ADPKD);
(4) History of kidney transplant or other solid organ transplant;
(5) Active systemic or specific organ infection (e.g., pneumonia,
osteomyelitis);
(6) Known hypersensitivity to mesenchymal stem cells or any component
of the cell culture medium;
(7) History of severe allergic or anaphylactic reactions to biological
products (e.g., blood-derived products);
(8) History of coagulation disorders (thromboembolism, pulmonary
embolism, deep vein thrombosis);
(9) History of malignancy or current malignant disease;
(10) Elevated tumor markers (AFP, CEA, CA199, CA125, etc.);
(11) Pregnant women or women planning pregnancy within 3 months
after MSC therapy;

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(12) Participation in drug-related clinical trials within the past 2 months;
(13) Any form of drug abuse, mental illness, or other conditions
considered by the investigator as potentially affecting the trial's validity
or the subject's health.
4.3 Study Termination Criteria
Criteria for subject withdrawal include:
(1) Occurrence of serious adverse events or complications related to MSC
injection or the CKD disease itself;
(2) Loss to follow-up or death of the subject;
(3) Voluntary request for withdrawal by the subject or their legal
guardian.
The withdrawal rate will be calculated at the end of the study. The
reason and date of withdrawal will be recorded in detail. For subjects who
withdraw from the study, appropriate treatment will be provided based on
the judgment of the investigators at the participating center, and
follow-up of their treatment outcomes may continue.
5. Methods and Technical Route
5.1 Interventions
All eligible patients will be randomly assigned to the experimental
group (MSC group) or the control group (placebo group), while
continuing to receive conventional therapy. The experimental group will
receive an intravenous injection of allogeneic MSCs at a dose of

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1.0*106/kg. The control group will receive a placebo, which is an
intravenous infusion of 250 mL of 0.9% sodium chloride injection,
matching the appearance and administration procedure of the MSC
infusion..
5.2 Sample Size Determination
This trial is an exploratory study. A total of 32 subjects are planned
to be enrolled, with 16 each for hypertensive nephropathy and diabetic
nephropathy, as a preliminary exploration of the treatment. For each
pathological type, subjects will be randomly allocated in a 1:1 ratio to the
experimental and control groups, meaning 8 subjects per group per
pathology (total experimental group n=16, control group n=16).
5.3 Randomization
Subjects who provide informed consent will be allocated in a 1:1
ratio to the experimental group or the control group. Computer-generated
randomization will be used to determine group assignment. A random
number sequence will be generated using SPSS 27.0 (or higher) software
(provided by a statistical professional). According to this sequence,
corresponding group codes will be placed into sealed, opaque envelopes.
Upon sequential enrollment of subjects, the group assignment will be
determined
by
the
code
inside
the
envelope,
administering
the
corresponding treatment.
5.4 Data Collection and Follow-up

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5.4.1 Information Collected
Table 1. Study Visit Schedule and Data Collection
Study Visit
creening/Baseline/Randomization
Post-treatment
Month 1, 3, 6, 12
Month 1,3,6,12 and
End of Study
Sign Informed
Consent
√
Medical History
√
√
√
Symptoms and
Signs
√
√
√
ECG or
Echocardiography
(if needed)
√
√
√
Routine
Examinations
√
√
√
Treatment
Modality
√
√
√
Concomitant
Medication
Record
√
√
√

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Note: All adverse events and side effects of the drug will be recorded.
Concomitant medications will be recorded and categorized by drug type.
5.4.2 Baseline Data
(1) Demographic information: age, gender, ethnicity, BMI, blood
pressure, smoking history, clinical diagnosis, etc.
(2) Laboratory tests:
* Renal and Liver Function: eGFR, serum creatinine, cystatin C, blood
urea nitrogen, serum albumin, cholesterol, triglycerides, LDL, HDL, uric
acid, blood glucose.
* Other System Functions: NT-proBNP, PTH, 1,25-(OH)2-Vitamin D3,
complement, T lymphocytes.
* Urinalysis: protein qualitative, urine red blood cell count.
* 24-hour urine protein quantification (mg/24h).
* Complete Blood Count: red blood cell count, hemoglobin.
* Tumor Markers (AFP, CEA, CA125, CA199, etc.).
(3) Baseline treatment regimen: use of antihypertensive, lipid-lowering,
Renal Function
Related
Parameters
√
√
√
Primary Endpoint
Events
√
√
√
Secondary
Endpoint Events
√
√
√

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hemostatic, and other relevant medications.
5.4.3 Follow-up Data
(1) Primary and secondary endpoints;
(2) Routine laboratory tests:
* Renal and Liver Function: eGFR, serum creatinine, cystatin C, blood
urea nitrogen, serum albumin, cholesterol, triglycerides, LDL, HDL, uric
acid, blood glucose.
* Other System Functions: NT-proBNP, PTH, 1,25-(OH)2-Vitamin D3,
complement, T lymphocytes.
* Urinalysis: protein qualitative, urine red blood cell count.
* 24-hour urine protein quantification (mg/24h).
* Complete Blood Count: red blood cell count, hemoglobin, etc.
(3) Imaging examinations: ECG, echocardiography, renal/renal vessel
ultrasound, chest CT, etc. (if necessary).
5.4.4 Strategies to Ensure Data Integrity
(1) Routine Clinical Visit Data Collection: This study will use paper
Case Report Forms (CRFs) for data collection. An electronic data capture
(EDC) system (or an Access database) will be used for data management.
Double data entry with comparison will be employed for data entry and
verification, followed by corresponding data checks and cleaning. During
the study period, the principal researchers will review the trial data
monthly to ensure internal consistency.

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(2) Subject Reminders: Automated reminder messages will be sent to
subjects in advance via WeChat group, SMS, or phone calls to minimize
sample loss due to loss to follow-up and/or withdrawal.
5.5 Technical Route Diagram

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6. Observation Items and Time Points
Primary Endpoints: The estimated glomerular filtration rate (eGFR)
and 24-hour urine protein quantification levels at 1, 3, 6, and 12 months
will serve as the primary outcome measures.
Secondary Endpoints: Changes in other renal function parameters
(urine
red
blood
cells,
cystatin
C,
blood
urea
nitrogen,
urine
protein/creatinine ratio, etc.), changes in other organ system function
parameters (ECG/echocardiography, complete blood count, liver function,
NT-proBNP,
parathyroid
hormone,
25-dihydroxyvitamin
D,
etc.),
immune-inflammatory markers (interleukins, CRP, T-cell subset analysis,
complement levels, etc.), and the incidence of adverse events at 1, 3, 6,
and 12 months will serve as secondary outcomes.
Severity will be graded according to the Common Terminology
Criteria for Adverse Events (CTCAE) v5.0:
Grade 1: Mild; asymptomatic or mild symptoms; clinical or diagnostic
observations only; intervention not indicated.
Grade 2: Moderate; minimal, local, or noninvasive intervention indicated;
limiting age-appropriate instrumental Activities of Daily Living (ADL).
Grade
3:
Severe
or
medically
significant
but
not
immediately
life-threatening;
hospitalization
or
prolongation
of
hospitalization
indicated; disabling; limiting self-care ADL.

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Grade 4: Life-threatening consequences; urgent intervention indicated.
Grade 5: Death related to adverse event.
7. Observation of Adverse Events
Potential adverse reactions associated with MSC therapy may include: (1)
Systemic immunogenic reactions to the injected cells, local inflammation,
and long-term immune responses; (2) Cell dysfunction, viral infection,
and post-injection infectious complications; (3) Fever, local pain; (4)
Pulmonary and renal thromboembolism, cardiac and hepatic fibrosis; (5)
although the mainstream view suggests that MSC therapy may inhibit
tumor occurrence and metastasis, there are reports indicating a potential
increased risk of tumor recurrence; (6) Xenogeneic contamination
reactions potentially leading to acute inflammatory responses, etc.
8. Quality Control and Quality Assurance
The investigators will be responsible for screening subjects according to
the inclusion/exclusion criteria, recording research data in the CRFs, and
entering the data into the electronic database. Simultaneously, they will
strictly adhere to the timeline and maintain close contact with the subjects
to avoid enrolling ineligible subjects, prevent subjects from voluntarily
withdrawing, ensure subjects receive the assigned treatment, and prevent
crossover between groups. A Data and Safety Monitoring Board (DSMB)
composed of two statisticians or clinicians experienced in clinical
research design and data analysis will be established. The DSMB will

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independently
monitor
the
study,
oversee
randomization,
data
management, safety monitoring, and statistical analysis, blinded to the
specific group assignments and treatments of the subjects. Once quality
assurance procedures are completed, the database will be locked.
9. Statistical Analysis
All statistical analyses will be performed based on the Intention-to-Treat
(ITT) principle, regardless of poor compliance, withdrawal, or loss to
follow-up. We will conduct independent observational analyses of the
two treatment regimens and report the trial results accurately. Continuous
variables conforming to a normal distribution will be expressed as mean
± standard deviation, and comparisons between two groups will use
T-tests, while comparisons among multiple groups will use analysis of
variance (ANOVA). Non-normally distributed continuous variables will
be expressed as median and interquartile range [M (P25, P75)].
Categorical variables will be expressed as percentages, and comparisons
between groups will use chi-square tests. Results will be reported as
Hazard Ratios (HR) with 95% Confidence Intervals (CI). The decline rate
of eGFR will be compared between groups using a linear mixed-effects
model.
Survival
analysis
will
employ
Cox
regression
models.
Considering the exploratory nature and purpose of this preliminary study,
the limited sample size, and the consequent relatively limited statistical
power, we will conduct objective preliminary comparative analyses and

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interpret the results cautiously. All statistical analyses will be performed
using SPSS 27.0 or R version 4.2.3 software.
10. Ethics of the Clinical Research
The clinical research will adhere to the World Medical Association's
Declaration of Helsinki and other relevant regulations. Before the study
begins, the protocol must be approved by the Institutional Review
Board/Ethics Committee. Prior to enrolling any subject in this study, the
investigator is responsible for providing the subject or their legal
representative with a complete and comprehensive explanation of the
study's purpose, procedures, potential benefits, and risks. Written
informed consent must be obtained. Subjects should understand that they
have the right to withdraw from the study at any time. The informed
consent forms will be retained as clinical research documents for future
reference. The privacy of subjects and the confidentiality of their data
will be protected throughout the research process.
11. Study Timeline
Jan. 1, 2024 – Jan. 31, 2025: Preliminary preparation, study protocol
design, investigator meeting, ethics committee review, trial registration.
Feb. 1, 2025 – Jul. 31, 2026: Trial initiation, patient enrollment and
follow-up, monitoring visits, data collection, management of blood
samples and adverse drug events.

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Aug. 1, 2026 – Aug. 31, 2027: Data consolidation, document archiving,
statistical analysis, manuscript writing and publication, reporting of
results.
12. References
[1] Kalantar-Zadeh K, Jafar T H, Nitsch D, et al. Chronic kidney disease[J]. The Lancet, 2021,
398(10302): 786-802.
[2] Feng T, Xu Y, Zheng J, et al. Prevalence of and risk factors for chronic kidney disease in ten
metropolitan areas of China: a cross-sectional study using three kidney damage markers[J]. Renal
Failure, 2023, 45(1).
[3] Bikbov B, Purcell C A, Levey A S, et al. Global, regional, and national burden of chronic
kidney disease, 1990–2017: a systematic analysis for the Global Burden of Disease Study 2017[J].
The Lancet, 2020, 395(10225): 709-733.
[4] Liyanage T, Ninomiya T, Jha V, et al. Worldwide access to treatment for end-stage kidney
disease: a systematic review[J]. Lancet, 2015, 385(9981): 1975-1982.
[5] Rodríguez-Fuentes D E, Fernández-Garza L E, Samia-Meza J A, et al. Mesenchymal Stem
Cells Current Clinical Applications: A Systematic Review[J]. Archives of Medical Research, 2021,
52(1): 93-101.
[6] Han Ying, Guo Changcun, Shi Yongquan. [Expert consensus on standardized stem cell
transplantation for the treatment of decompensated liver cirrhosis (2021)][J].
[7] Chun S, Choi C, Kim M S, et al. Safety and tolerability of bone marrow–derived
mesenchymal stem cells in lupus animal models and a phase I clinical trial in humans[J]. Lupus,
2022, 31(10): 1245-1253.
[8] Ranjbar A, Hassanzadeh H, Jahandoust F, et al. Allogeneic adipose-derived mesenchymal
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Current Research in Translational Medicine, 2022, 70(2): 103324.
[9] Packham D K, Fraser I R, Kerr P G, et al. Allogeneic Mesenchymal Precursor Cells (MPC) in
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A
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Dose
Escalation
Study[J].
EBioMedicine, 2016, 12: 263-269.
[10] Hickson L J, Abedalqader T, Ben-Bernard G, et al. A Systematic Review and Meta-Analysis
of Cell-Based Interventions in Experimental Diabetic Kidney Disease[J]. Stem Cells Translational
Medicine, 2021, 10(9): 1304-1319.
[11] Wang Y, Shan S, Guo B, et al. The Multi-Therapeutic Role of MSCs in Diabetic
Nephropathy[J]. Frontiers in Endocrinology, 2021, 12.
[12] Swaminathan M, Stafford-Smith M, Chertow G M, et al. Allogeneic Mesenchymal Stem
Cells for Treatment of AKI after Cardiac Surgery[J]. Journal of the American Society of
Nephrology, 2018, 29(1): 260-267.
[13] Makhlough A, Shekarchian S, Moghadasali R, et al. Bone marrow–mesenchymal stromal cell
infusion in patients with chronic kidney disease: A safety study with 18 months of follow-up[J].
Cytotherapy, 2018, 20(5): 660-669.

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