移植人脐带干细胞改善乙型肝炎失代偿肝硬化患者的肝功能

doi:10.3969/j.issn.2095-4344.0922

中国组织工程研究 ›› 2018, Vol. 22 ›› Issue (25): 3993-4000.doi: 10.3969/j.issn.2095-4344.0922

• 干细胞移植 stem cell transplantation • 上一篇下一篇

移植人脐带干细胞改善乙型肝炎失代偿肝硬化患者的肝功能

傅青春^1，2，金银鹏^1，2，王晓今¹，李莉²，王皙¹，李洪超¹，汪照静³，周丰¹，臧祖胜¹，施莉琴¹，李震宇¹，陈成伟¹

¹解放军南京军区上海肝病研究中心，上海市 200235；²复旦大学附属上海市公共卫生临床中心，上海市 201508；³深圳北科细胞工程研究所，广东省深圳市 518040

修回日期:2018-04-05 出版日期:2018-09-08 发布日期:2018-09-08
通讯作者: 陈成伟，解放军南京军区上海肝病研究中心，上海市 200235
作者简介:傅青春，男，汉族，1986年第二军医大学毕业，硕士，主任医师，主要从事干细胞研究。并列第一作者：金银鹏，男，汉族，硕士，从事干细胞研究。
基金资助:
南京军区医学创新课题(10MA001)

Transfusion with human umbilical cord mesenchymal stem cells improves liver function in hepatitis B patients with decompensated liver cirrhosis

Fu Qing-chun^{1, 2}, Jin Yin-peng^{1, 2}, Wang Xiao-jin¹, Li Li², Wang Xi¹, Li Hong-chao¹, Wang Zhao-jing³, Zhou Feng¹, Zang Zu-sheng¹, Shi Li-qin¹, Li Zhen-yu¹, Chen Cheng-wei¹

¹Shanghai Liver Disease Research Center, Nanjing Military Command, Shanghai 200235, China; ²Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508, China; ³Shenzhen Beike Cell Engineering Research Institute, Shenzhen 518040, Guangdong Province, China

Revised:2018-04-05 Online:2018-09-08 Published:2018-09-08
Contact: Chen Cheng-wei, Shanghai Liver Disease Research Center, Nanjing Military Command, Shanghai 200235, China
About author:Fu Qing-chun, Master, Chief physician, Shanghai Liver Disease Research Center, Nanjing Military Command, Shanghai 200235, China; Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508, China. Jin Yin-peng, Master, Shanghai Liver Disease Research Center, Nanjing Military Command, Shanghai 200235, China; Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508, China. Fu Qing-chun and Jin Yin-peng contributed equally to this work.
Supported by:
the Medical Innovation Project of Nanjing Military Command, No. 10MA001

摘要/Abstract

摘要：

文章快速阅读：

文题释义：
失代偿肝硬化：肝硬化是由一种或多种病因长期、反复刺激造成肝脏弥漫性损害，其特点为慢性弥漫性结缔组织增生，肝细胞变性坏死、再生和肝小叶结构损害及假小叶形成。早期肝功能尚可代偿，肝硬化发展到一定程度，超出肝功能的代偿能力，即称为失代偿肝硬化。
脐带间充质干细胞：是指存在于新生儿脐带组织中的一种多功能干细胞，它能分化成许多种组织细胞，具有广阔应用前景。

摘要
背景：失代偿肝硬化是危机生命的慢性肝疾病并发症，是肝移植的主要指标之一。
目的：探索人脐带间充质干细胞治疗乙肝肝硬化失代偿患者的安全性和有效性。
方法：间充质干细胞治疗组包括17例乙肝肝硬化失代偿患者及对照组包含17例匹配的乙肝失代偿患者。所有患者接受标准乙肝肝硬化治疗，包括口服抗病毒核酸类似物。间充质干细胞治疗组接受肘静脉注射人脐带间充质干细胞。
结果与结论：间充质干细胞移植患者肝功能有所改善，白蛋白、胆红素、乳酸脱氢酶及碱性磷酸酶水平有所恢复，且患者Child-Pugh及MELD评分明显恢复。表明人脐带间充质干细胞静脉注射入乙肝肝硬化失代偿患者体内可改善肝功能。

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程
ORCID: 0000-0002-7217-5315 (Fu Qing-chun)

关键词: 失代偿肝硬化, 人脐带间充质干细胞, 移植, 肝功能, 安全性, 乙肝肝硬化, 标准治疗, 干细胞

Abstract:

BACKGROUND: Decompensated liver cirrhosis, a life-threatening complication of chronic liver disease, is one of the major indications for liver transplantation.
OBJECTIVE: To evaluate the safety and efficacy of human umbilical cord mesenchymal stem cell (hUC-MSC) transplantation in hepatitis B patients as a treatment for decompensated liver cirrhosis.
METHODS: The mesenchymal stem cell (MSC) treatment group contained 17 hepatitis B patients with decompensated liver cirrhosis and the control group contained 17 matched controls. All patients received conventional treatment for hepatitis B cirrhosis, including the oral administration of antiviral nucleoside analogues. Patients in the MSC treatment group received hUC-MSC infusions into the elbow vein, while control patients received no injection. Follow-up examinations were conducted after treatment to evaluate clinical symptoms, liver function and blood coagulation function. In addition, the severity of cirrhosis was assessed based on the Child-Pugh and Model for End-Stage Liver Disease scores.
RESULTS AND CONCLUSION: In this study, we revealed that hUC-MSC-treated patients had an improvement in liver function, as indicated by the recovered albumin, bilirubin, lactate dehydrogenase and alkaline phosphatase levels to some extent. Additionally, hUC-MSC-treated patients had better Child-Pugh and Model for End Stage Liver Disease scores than control patients. Therefore, our findings reveal that the transfusion with hUC-MSCs improves the liver function, which represents a safe approach for treating decompensated liver cirrhosis in hepatitis B patients.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Liver Cirrhosis, Umbilical Cord, Tissue Engineering

中图分类号:

R394.2

傅青春，金银鹏，王晓今，李莉，王皙，李洪超，汪照静，周丰，臧祖胜，施莉琴，李震宇，陈成伟. 移植人脐带干细胞改善乙型肝炎失代偿肝硬化患者的肝功能[J]. 中国组织工程研究, 2018, 22(25): 3993-4000.

Fu Qing-chun, Jin Yin-peng, Wang Xiao-jin, Li Li, Wang Xi, Li Hong-chao, Wang Zhao-jing, Zhou Feng, Zang Zu-sheng, Shi Li-qin, Li Zhen-yu, Chen Cheng-wei. Transfusion with human umbilical cord mesenchymal stem cells improves liver function in hepatitis B patients with decompensated liver cirrhosis[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(25): 3993-4000.

图/表（结果） 2

Quantitative analysis of patients

The study flow is presented in Figure 1. A total of 138 hepatitis B patients with DLC were recruited. Three patients were excluded because they did not meet the inclusion criteria, and 101 patients were excluded because they did not provide one-on-one correspondence. Seventeen hepatitis B patients were recruited to the human umbilical cord-derived mesenchymal stem cell (hUC-MSC) treatment group and the control group contained 17 clinically matched individuals.

Baseline comparisons and clinical symptoms

The baseline characteristics are presented in Table 1. The patients in the MSC-treated and control groups were similar regarding sex, age and biochemical data. The baseline Child-Pugh (P=0.149) and MELD (P=0.661) scores were also similar. The patients in the two groups did not differ regarding the course of DLC (P=0.333; Figure 2A).

and control groups had undetectable levels of HBV DNA in their serum, with no significant differences detected between the groups (P=0.247). The antiviral length was also comparable between the groups (P=0.704). Ascites gradually decreased in both the MSC-treatment and control groups following the administration of albumin and diuretics and no significant differences were detected between the groups (P=0.203).

Liver function tests

Forty-eight weeks after treatment, Alb levels increased from (28.9±2.8) g/L to (33.9±5.7) g/L in the MSC-treatment group, which was significantly higher than the baseline (P < 0.05; Figure 2A). PAB increased from (62.1±32.7) g/L to (98.7±41.6) g/L in MSC-treated patients, which was also significantly higher than the baseline value (P < 0.05; Figure 2B). Alb and PAB levels did not differ significantly to the baseline values after 48 weeks of conventional treatment alone (Figure 2A, B). The differences in Alb and PAB levels were not significant between the two groups at different time points (Figure 2A, B).

TB levels at 48 weeks after treatment were: MSC-treatment group, 53.9 to 37.0 μmol/L; control group, (54.6±45.3) to (55.7±47.0) μmol/L (Figure 2C, D). The differences between the two groups at each time point were not statistically significant.

The levels of LDH and AKP were significantly lower in the MSC-treatment group compared to the control group 48 weeks after treatment (P < 0.05; Figure 2E, F). Forty-eight weeks after treatment, the LDH values were: MSC-treatment group, (191.4±63.7) U/L; control group, (272.2±105.3) U/L. The AKP values 48 weeks after treatment were: MSC-treatment group, (87.2±33.7) U/L; control group, (133.8±58.3) U/L. The levels of ALT, γ-GT and CPK did not differ significantly between the MSC-treatment and control groups at all time points (Figure 2G, H, I).

Blood coagulation tests

Cirrhosis patients have an increased risk for bleeding. We assessed the coagulation function in HBV patients with DLC by measuring the PT and APTT. The PT did not differ significantly between the MSC-treated and control patients at various time points after treatment (Figure 3A). We observed a faster APTT in the MSC-treatment group 4 weeks after treatment and the APTT continued to decrease in the MSC-treated patients until 48 weeks after treatment (Figure 3B). The APPT values at 48 weeks after treatment were: MSC-treatment group, 39.9 seconds (7.5-second decrease); control group, 43.1 seconds (7.5-second increase). The differences between the two groups at each time point were not statistically significant.

Renal function

Cr levels in the blood were measured to assess renal function^[35]. We detected a decline in Cr levels in the blood of MSC-treated patients compared to control patients (Figure 3C), which indicated improved renal function. However these differences were not statistically significant, although this may reflect the low number of participants, rather than a genuine lack of significance.

Child-Pugh and MELD scores

The Child-Pugh and MELD scores were calculated to evaluate the severity of cirrhosis in the patients^[30]. The Child-Pugh score declined significantly from 9.4 points at the baseline to 7.5 points measured 48 weeks after treatment in the MSC-treatment group, (P < 0.05; Figure 3D), indicating a better prognosis. The MELD score also declined from 15.8 to 13.3 points in MSC-treated patients, while in the control group, the MELD score rose slightly from 15.1 points at the baseline to 15.5 points 48 weeks after treatment (Figure 3E). This indicates a better prognosis in patients treated with hUC-MSCs compared to patients who received conventional treatment alone, although the differences between the two groups in each time point were not statistically significant.

Evaluation of potential complications following hUC-MSC transfusion

No abnormal changes were detected in the vital signs, electrocardiogram results and renal function of patients following transfusion with hUC-MSCs which were observed at the time of transfusion. Two patients developed a fever and a reduced platelet count was detected in two further patients. In one patient we observed an increase in the INR to 8.21, which recovered baseline after 72 hours without any treatment, although this decreased within 4 days and no hemorrhage-related complications were observed. No tumor development was observed during follow-up examinations in MSC-treated patients. Other potential serious adverse reactions associated with end-stage liver diseases, such as hepatorenal syndrome, hepatic encephalopathy and pleural effusion, were not related to MSC-treatment. There was no statistically significant difference in the adversity of the reaction between the two groups.

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引言

Hepatitis B virus (HBV) is a major cause of cirrhosis and approximately one million people die each year from cirrhosis-related complications as a result of HBV infection^[1]. HBV activates hepatic stellate cells in the liver, causing the deposition of a collagen matrix. The progression of fibrosis gradually disrupts the histological and vascular architecture of the liver, leading to impaired liver function and portal hypertension^[2]. The severity of cirrhosis can be monitored by specific hepatocyte function tests that measure the production of specific markers^[3-5].

Progression to decompensated liver cirrhosis (DLC) has a significantly negative impact on the disease prognosis, with an estimated 5-year survival rate of just 14%^[6]. In many cases, liver transplantation represents the best treatment for HBV-related DLC, but it is significantly restricted by the lack of donors^[7-9]. Liver cell transplantation is now emerging as an alternative to the whole organ transplantation^[10-12]. Mesenchymal stem cells (MSCs) are multipotent, capable of self-renewal and multi-directional differentiation. Additionally, they are reported to have anti-inflammatory and immune-modulating functions and can improve tissue vascularization^[13-18]. These characteristics have been exploited in a wave of attempts to treat a multitude of diseases by MSC transplantation^[19-20].

Human umbilical cord mesenchymal stem cells (hUC-MSCs) represent a useful source of MSCs for transplantation, as they avoid the ethical concerns associated with the use of stem cells cultured from embryonic tissue and the limitations of stem cells cultivated from adult differentiated tissue^[21-24]. hUC-MSCs are easy to cultivate and have been shown to differentiate along a hepatic lineage in vivo. Hepatocyte-like cells differentiated from hUC-MSCs can express hepatic markers, store glycogen and produce urea^[25-28]. These findings suggest that hUC-MSCs may be suitable for liver cell transplantation to treat DLC in HBV patients. In this study, we have transplanted hUC-MSCs into patients with decompensated HBV cirrhosis via peripheral intravenous infusion and evaluated the efficacy of this treatment in improving the clinical outcome. We monitored the patients’ quality of life and clinical symptoms, particularly liver function, during one year of follow-up examinations.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

材料方法

Design

Historical control study.

Time and setting

The experiment was performed in the 85^th Hospital of Chinese PLA between November 2010 and May 2011.

Subjects

This study was registered with the ClinicalTrial.org (registration No. NCT01342250). Ethical approval was obtained from the 85^th Hospital of Chinese PLA with the approval No. [2013] Ethical No. 18, and all patients provided written informed consent prior to the beginning of the study. Seventeen patients with chronic HBV and diagnosed with DLC and ascites according to theirs history of disease, laboratory test results and imaging results were recruited from the Shanghai Clinical Liver Disease Research Center of Nanjing Military Region between November 2010 and May 2011. A total of 138 hepatitis B patients with DLC were recruited. Three patients were excluded because they did not meet the inclusion criteria, and 101 patients were excluded because they did not provide one-on-one correspondence. These patients comprised the MSC-treatment group containing 17 hepatitis B patients with DLC (12 males and 5 females; mean age, 56.2 years) and the control group containing 17 clinically matched individuals (11 males and 6 females; mean age, 56.9 years).

The inclusion criteria were as follows: (1) informed consent was provided and treatment received voluntarily; (2) 18-70 years of age; (3) clinical diagnosis of decompensated HBV cirrhosis^[29], Child-Pugh score^[30] of 7-12, Model for End-Stage Liver Disease (MELD) score^[30] of ≤ 21; (4) life expectancy > 3 months; (5) cirrhosis treated with antiviral agents^[29-30]. The exclusion criteria were: (1) drug allergies or an allergic constitution; (2) severe diseases of the cardiovascular, respiratory or endocrine system, or of the blood; (3) severe infection; (4) diagnosed or suspected malignant tumor; (5) brain damage caused by a severe neurological deficit; (6) hepatic encephalopathy and/or hepatorenal syndrome; (7) spontaneous peritonitis within the last two weeks or varices bleeding within the last 4 weeks; (8) HIV positive; (9) liver transplant planned in the next three months; and (10) pregnancy, possibility of pregnancy or breastfeeding.

Methods

Preparation and identification of hUC-MSCs

The toxicity of hUC-MSCs was extensively tested in vitro and in animal models (unpressed). Acute toxicity was tested in mice, long-term toxicity was tested in monkeys and local irritation was tested inguinea pigs. The potential for malignant transformation (soft agar assay) and cell proliferation were analyzed in vitro^[31]. These trials were implemented in qualified laboratories.

The preparation of stem cells was approved by the China National Accreditation Service for Conformity Assessment (CNAS), the ISO 9001 Quality Management Systems Requirements (certification number: IS00110Q2149ROS/4403), the American Association of Blood banks (AABB) and the International Society For Cellular Therapy (ISCT). Quality control was very strictly implemented and each stem cell preparation was given a barcode, by which the source of infused stem cells can be easily traced. Flow cytometry test results of the present study were in accordance with the appraised standards released by the ISCT for hUC-MSCs^[32]. The expression levels of various markers (CD29⁺/CD44⁺/CD73⁺/CD90⁺/ CD105⁺/CD14⁺/CD19⁺/ CD34⁺/CD45⁺/HLA-DR) confirmed that hUC-MSCs are capable of multi-directional differentiation.

Treatment and transfusion of hUC-MSCs

It is standard practice to provide antiviral therapy to patients with decompensated HBV cirrhosis to slow the progression of the disease and reduce the need for liver transplantation^[33]. We administered antiviral nucleoside analogues orally and on demand. The purpose of this treatment was to inhibit the replication of HBV DNA. All fundamental situations of patients were similar. In the MSC-treatment group, lamivudine (LAM; Shanghai GSK) was administered to three patients, adefovir dipivoxil (Shanghai Chiatai tianqing) to four patients, entecavir (Shanghai Chiatai tianqing) to eight patients and a combination of LAM and entecavir was administered to two patients. In the control group, seven patients received LAM, two patients received adefovir dipivoxil, two patients received entecavor and six patients received a combination therapy of LAM and entecavir. Ascites was managed by infusion with albumin and diuretics. Patients in the MSC-treatment group received three transfusions of (0.5-2)×10⁸ hUC-MSCs which were suspended in 100 mL of 0.9% normal saline continuously through the right elbow vein, with 3 days between transfusions in addition to conventional treatment. Patients in the control group received 100 mL of 0.9% normal saline in addition to conventional treatment.

Follow-up

Following treatment with hUC-MSCs, the quality of life, clinical symptoms and any signs of change were monitored in all patients. Follow-up assessments were made at 1, 4, 12, 24, 36 and 48 weeks after treatment. The presence of HBV DNA in the serum usually indicates continuing viral replication^[33]. The degree of HBV infection was determined by measuring the viral load — specifically the quantification of HBV DNA in the serum of participants. As a measure of successful disease management, we monitored that the antiviral length of time that HBV DNA levels were undetectable in the patient serum^[33]. The course of DLC can be monitored by measuring certain blood parameters; the serum levels of albumin (Alb), prealbumin (PAB), total bilirubin (TB), direct bilirubin (DB), lactate dehydrogenase (LDH), alanine aminotransferase (ALT), alkaline phosphatase (AKP), γ-glutamyltransferase (γ-GT), creatine phosphate kinase (CPK), prothrombin time (PT), creatinine (Cr) and activated partial thromboplastin time (APTT) were measured using an ELISA kit (Sigma, Santa Louis, MO, USA). The Child-Pugh and MELD scores were measured to assess the severity of the cirrhosis and to monitor any adverse reactions. These scores were determined as previously described^[34].

Main outcome measures

Liver function was assessed in this study.

Statistical analysis

Statistical analysis was performed using SPSS 13.0 for Windows software (SPSS, Chicago, IL, USA). All data are presented as the mean±SD. The baseline characteristics (sex, Child-Pugh score, phase of DCL, viral load, antiviral length and ascites) were compared between groups using the Pearson chi-square test. Baseline laboratory tests were compared between groups using the independent samples t-test. The liver function, blood coagulation function and renal function was detected by biochemical tests, such as Alb, PAB, TB, LDH, AKP, ALT, γ-GT, CPK, PT, APTT and Cr. The Child-Pugh and MELD scores were calculated with respective corresponding scale. Data were compared before and after treatment using the t-test. Comparison of data between groups after treatment was performed using the Mann-Whitney U test. A value of P < 0.05 was inferred statistically significant.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

讨论

In recent years, the transfusion of MSCs has improved the symptoms of liver disease in both animal models and human patients. hUC-MSCs represent a useful source of MSCs for transplantation as they are easier to harvest, pose minimal ethical implications and can be safely transplanted ^[36-37]. In order to validate hUC-MSCs as a standard treatment for HBV DLC, more clinical trials are required in human patients^[38-41]. In this study, we evaluated the efficacy of hUC-MSC transfusion as a treatment for DLC in HBV patients by evaluating liver function and patient well-being after treatment.

During follow-up examinations, MSC-treated patients reported a larger improvement in spirit, sleep and fatigue symptoms compared to control patients. This may reflect a reduction in toxins in the bloodstream that can affect the brain. Alternatively, these positive effects on well-being may be a side-effect of improved clinical symptoms. Whether MSCs secrete a factor that can adjust the mental state remains to be elucidated.

We also observed better results in tests of liver function in MSC-treated patients compared to saline-treated controls, including Alb, PAB, TB, DB, LDH and AKP levels. An improved liver function in MSC-treated patients was confirmed by higher Child-Pugh scores in these individuals compared to controls. Transplantation of hUC-MSCs may influence liver function through a variety of mechanisms; hUC-MSCs have been shown to differentiate along a hepatocyte lineage^[22] therefore may replace lost or damaged hepatocytes in the diseased liver. hUC-MSCs can be programmed to differentiate down a hepatocyte lineage by locally produced tissue specific factors, or by the combination of genetic material after fusion with adjacent cells^[42-44]. Transplanted MSCs have also been shown to have an anti-inflammatory effect, suppress the immune response, limit apoptosis in diseased tissue and promote vascularization^[13-18]; these factors would all contribute to the regeneration of damaged liver tissue^[42-44].

It has been shown that transplanted MSCs can reduce the symptoms of liver fibrosis in mice by producing matrix metalloproteinases, which degrade the extracellular matrix and inhibit collagen deposition to reduce matrix secretion by activated hepatic stellate cells ^[45]. Additionally, MSCs may either induce the apoptosis of activated hepatic stellate cells, or limit their activation to reduce collagen deposition^[45-48]. Portal hypertension could, in theory, be reduced by these anti-fibrotic effects, which may reduce the severity of ascites in patients with decompensated HBV cirrhosis. Although we did not observe any improvement in ascites in our patients, transfusion of hUC-MSCs into HBV DLC patients has been shown to reduce ascites in a previous study ^[49]. The lack of improvement in ascites following hUC-MSC transfusion in our study may be explained by the fact that an insufficient number of MSCs migrated to the liver, or differentiated into hepatocytes to have a positive clinical effect^[50-51]. We did not perform biopsies of liver tissue in this, or previous clinical trials, due to ethical issues, therefore were not able to confirm the migration of injected hUC-MSCs to the liver or investigate immunological changes after MSCs migrated to the liver. However, we have previously tested the therapeutic potential of cultured human adipose-derived MSCs in the liver using a rat model. We found that MSCs can migrate into the liver tissue through the pulmonary capillary network after injection. Additionally, we confirmed the ability of human adipose-derived MSCs to secrete hepatocyte growth factor and vascular endothelial growth factor in culture and promote the regeneration of damaged liver tissue through paracrine functions^[52]. These aspects should be investigated in human patients in future studies.

We did not observe any adverse effects following the transfusion of hUC-MSCs into the peripheral vein. Measurements of HBV DNA remained undetectable in the serum and we found no evidence of tumor development, despite previous reports that MSCs can promote tumorigenesis in response to chemotactic inflammation at sites of tumor formation ^[53]. In agreement with our findings there are no existing reports of tumor formation in clinical trials and MSC transfusion in combination with conventional treatments is considered a safe approach to the treatment of DLC^[45]. Despite this, the most suitable course of MSC treatment still needs to be determined for decompensated HBV cirrhosis.

The MSC dose and number of transplantations needs to be optimized and the timing of MSC migration to the liver after injection needs to be explored. Some authors have reported that the transplantation of too many cells or too many infusions can induce an immune response or cause complications such as pulmonary embolism^[54-55]. Methodological factors affecting the preparation and culture of MSCs, such as culture medium, oxygen tension, thawing, tissue source, genetic modification and the MSC survival time in the human body, all have a significant influence on the safety and effectiveness of MSC therapy^[19]. Different implantation sites such as the hepatic artery and portal vein and different cell numbers will be tested in the future, to try and optimize these conditions. Multicenter randomized controlled trials, analyzing a larger number of patients together with an extended follow-up time are also necessary to investigate the impact of MSC-treatment on advanced liver disease.

In conclusion, we have demonstrated that the transfusion of hUC-MSCs significantly improves the liver function and the well-being of HBV patients with DLC compared to conventional treatments alone.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

移植人脐带干细胞改善乙型肝炎失代偿肝硬化患者的肝功能

Transfusion with human umbilical cord mesenchymal stem cells improves liver function in hepatitis B patients with decompensated liver cirrhosis

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