Safety and feasibility of autologous bone marrow mesenchymal stem cells in treating chronic allograft nephropathy

doi:10.3969/j.issn.2095-4344.2014.32.010

Abstract

Abstract:

BACKGROUND: Chronic allograft nephropathy is a complication of kidney transplantation and most of patients will eventually develop transplant kidney dysfunction. Bone marrow mesenchymal stem cells as a low immunogenicity special cell population have been shown to have differentiation, transdifferentiation, paracrine and other basic functions, which have been successful used in other clinical areas. Based on this characteristic, bone marrow mesenchymal stem cells may play a therapeutic role in chronic allograft nephropathy.
OBJECTIVE: To study the safety and feasibility of autologus bone marrow mesenchymal stem cells transplantation via renal artery infusion and subsequent intravenous infusion guided by the digital subtraction angiography in the treatment of chronic allograft nephropathy.
METHODS: Eleven patients with chronic allograft nephropathy who were confirmed from March 2011 to January 2013 were enrolled, and then received transplant renal artery infusion once guided by the digital subtraction angiography and subsequent intravenous infusion twice of bone marrow mesechymal stem cells. Changes in serum creatinine, blood urea nitrogen, creatinine clearance, cystatin C, 24-hour urine protein, and β2 microglobulin in the blood and urinary were monitored in patients up to 1 year after treatment.
RESULTS AND CONCLUSION: Bleeding, transplant renal artery embolization, pseudoaneurysm and other related complications were not found in any of the 11 patients. The levels of serum creatinine, blood urea nitrogen and cystatin C were significantly decreased at 1 week and 1 month after cell therapy (P < 0.05), while after 3 months of treatment, there was no difference before and after treatment (P > 0.05). The creatinine clearance at 1 week and 1 month after treatment showed a remarkable increase, which were significantly different from that before treatment (P < 0.05), but after 3 months of treatment, the difference was not significant (P > 0.05). The level of 24-hour urine protein was significantly decreased after 7 days of treatment (P < 0.05), and no difference was found after 1 month (P > 0.05). The level of β2 microglobulin in the blood and urinary had no changes before and after treatment. These findings indicate that guided by the digital subtraction angiography, bone marrow mesenchymal stem cells via the renal artery infusion and subsequent intravenous infusion can improve kidney function of patients, but the cell dosage and infusion method remain to be solved.

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

全文链接：

Key words: bone marrow, mesenchymal stem cells, kidney transplantation, postoperative complications, nephrosis, kidney function tests

CLC Number:

R394.2

Zhang Lei, Chen Zheng1, Xie Si-sheng, Ma Jun-jie, Fang Jia-li, Li Guang-hui, Xu Lu, Zhang Yi-rui, Guo Yu-he, Pan Guang-hui. Safety and feasibility of autologous bone marrow mesenchymal stem cells in treating chronic allograft nephropathy[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(32): 5140-5145.

Figures/Tables 1

2.1 骨髓间充质干细胞的形态采用密度梯度离心法与贴壁筛选法相结合可以有效分离、纯化人骨髓间充质干细胞。细胞初接种入培养瓶时为圆形的单个核细胞，大小不一，与周围的血细胞相互混杂(图1A)。培养24 h后细胞开始出现贴壁，48-72 h细胞贴壁逐渐增多，72 h首次换液后，可见少量分散的短梭形及不规则多角形的初始贴壁细胞(图1B)。第4-8天细胞生长快，呈集落样生长，可见有粗大的突起伸出，细胞形态呈梭形。第10-12天后细胞克隆进一步扩大，呈大的长梭形，培养至第14-16天时贴壁细胞可达80%-90%融合，大量贴壁细胞呈长梭状成纤维细胞样形态并呈漩涡状排列(图1C，D)。 2.2 骨髓间充质干细胞的鉴定利用间充质干细胞贴壁的特性进行分离，传至第5代行成骨及成脂诱导，并行表面标记物测定。加入成骨诱导液21 d后，经茜素红染色，光镜下可见细胞内有红色钙结节沉积，加入成脂诱导液 14 d后，经油红O染色，光镜下可见细胞内出现脂肪颗粒(图2)。流式细胞术检测人骨髓间充质干细胞的表型为CD44+CD29+CD105+CD48-CD34-。 2.3 移植肾功能临床指标变化 2.3.1 治疗前后血肌酐变化情况骨髓间充质干细胞治疗后7 d、1个月时患者血肌酐值较治疗前下降，其差异有显著性意义(P < 0.05)。治疗3个月后血肌酐值与治疗前相比差异无显著性意义(P > 0.05)，见表1。 2.3.2 治疗前后尿素氮变化情况骨髓间充质干细胞治疗后7 d、1个月患者尿素氮水平较治疗前下降，其差异有显著性意义(P < 0.05)。治疗3个月后尿素氮水平与治疗前相比差异无显著性意义(P > 0.05)，见表1。 2.3.3 治疗前后内生肌酐清除率变化情况骨髓间充质干细胞治疗后7 d、1个月患者内生肌酐清除率较治疗前升高，其差异有显著性意义(P < 0.05)。治疗3个月后内生肌酐清除率与治疗前相比差异无显著性意义(P > 0.05)，见表1。 2.3.4 治疗前后胱抑素C变化情况骨髓间充质干细胞治疗后7 d、1个月患者胱抑素C水平较治疗前下降，其差异有显著性意义(P < 0.05)。在骨髓间充质干细胞治疗3个月后胱抑素C水平与治疗前相比差异无显著性意义(P > 0.05)，见表1。 2.3.5 治疗前后24 h尿蛋白变化情况骨髓间充质干细胞治疗后7 d患者24 h尿蛋白水平较治疗前下降，其差异有显著性意义(P < 0.05)。治疗1个月后24 h尿蛋白水平与治疗前相比差异无显著性意义(P > 0.05)，见表1。 2.3.6 治疗前后血β2微球蛋白变化情况骨髓间充质干细胞治疗后患者血β2微球蛋白水平与治疗前相比差异无显著性意义(P > 0.05)，见表1。 2.3.7 治疗前后尿β2微球蛋白变化情况骨髓间充质干细胞治疗后患者尿β2微球蛋白水平与治疗前相比差异无显著性意义(P > 0.05)，见表1。 2.4 不良事件及随访情况 11例患者中，1例患者在第1次骨髓间充质干细胞输注后3 d出现发热、咳嗽、咳痰等，血培养及痰培养均未见细菌、真菌生长。1例患者在第2次骨髓间充质干细胞输注过程中出现浑身瘙痒，皮肤风团等过敏症状，予抗过敏治疗后治愈，上述2例患者骨髓间充质干细胞稀释液培养后未见细菌、真菌生长。未见出血、动脉栓塞、假性动脉瘤等并发症。所有患者首次输注骨髓间充质干细胞后12个月时行移植肾彩超检查，未见移植肾血流分布特殊异常，未见假性动脉瘤等。"

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