Human urine-derived stem cell transplantation for the treatment of chronic kidney disease in rats

doi:10.3969/j.issn.2095-4344.2016.32.019

Abstract

Abstract:

BACKGROUND: Urine-derived stem cells are most likely to come from the kidney tissue, and therefore, these cells are more adaptable to kidney microenvironment, providing a new option for the treatment of kidney diseases.
OBJECTIVE: To explore the therapeutic efficacy of human urine-derived stem cells on chronic nephropathy rats.
METHODS: The fresh urine samples of healthy people were collected, and then human urine-derived stem cells were extracted and cultured in vitro. Twenty Sprague-Dawley rats were used to prepare chronic nephropathy models, and given injection of human urine-derived stem cell suspension (experimental) or normal saline (control) into the renal cortex, respectively. Another 10 healthy rats were used as controls. Therapeutic effects on renal function were assessed by detection of serum creatinine level and glomerular filtration rate in the three groups. The kidney tissues of rats were taken and observed histomorphologically in each group.
RESULTS AND CONCLUSION: Human urine-derived stem cells were found to remarkable improve rat’s renal function as well as reduce the histomorphological changes in the kidney tissues of rats. Compared with the control group, the serum creatinine level was decreased while the glomerular filtration rate was increased significantly in the experimental group; CD68 expression and infiltration of interstitial inflammatory cells were also markedly reduced in the experimental group. To conclude, human urine-derived stem cells can improve the renal function of chronic nephropathy rats.

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: Nephrosis, Urine, Stem Cell Transplantation, Tissue Engineering

CLC Number:

R394.2

Zhao Ya-pei, Liu Cui-jing, Yang Cui-ying, Fan Shi-ying, Zhang Li-jian, Zhao Yan-qin, Xie Ying-xin. Human urine-derived stem cell transplantation for the treatment of chronic kidney disease in rats[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(32): 4838-4844.

Figures/Tables 2

2.1 实验动物数量分析实验共用大鼠30只，全部进入结果分析，无脱失。 2.2 人尿源干细胞形态及表面标志尿源干细胞形状小而饱满，增殖迅速，生长旺盛，培养1周细胞呈纺锤形，见图1。流式细胞仪检测人尿源干细胞表面标记物 CD29、CD146、CD44、CD90、CD24、CD73、CD105呈阳性表达，CD45、CD34、CD31等造血细胞表面抗原呈阴性表达(表1，图2)，符合间充质干细胞的定义[12]。 2.3 大鼠肾功能学监测结果分析由表2可见，3组SD大鼠血清肌酐水平差异有显著性意义(P < 0.05)。实验组大鼠在尿源干细胞移植后2周内，其血清肌酐水平下降迅速，4周后血清肌酐水平慢慢趋于平稳，与对照组相比差异有显著性意义(P < 0.05)；实验组大鼠血清肌酐虽有所下降，但尚未恢复到正常水平，与正常组比较差异有显著性意义(P < 0.05)。对照组大鼠血清肌酐水平有持续明显升高状态，与正常组、实验组间存在明显的差异性。由表3可见，3组SD大鼠肾小球滤过率的变化趋势与血清肌酐类似，实验组大鼠肾小球滤过率的提高主要发生在注射后的4周内，实验组肾小球滤过率与对照组、正常比较，差异有显著性意义(P < 0.05，P < 0.01)，此后各组大鼠肾小球滤过率水平均趋于平稳，没有明显的变化。 2.4 大鼠肾脏切片苏木精-伊红染色从图3可见，正常组大鼠肾脏肾小球结构正常，间距、大小较为均匀，肾脏皮质中肾小管排列紧密、结构正常，肾脏组织中无纤维化形成；对照组注射等量生理盐水大鼠肾脏肾小球数量减少、萎缩，毛细血管分叶，肾脏皮质中肾小管管腔增大，肾脏组织中有大量纤维增生；注射尿源干细胞实验组大鼠肾脏结构位于正常组与对照组之间，与对照组相比，实验组大鼠肾小球数量增多、形态尚规则，纤维组织增生明显减少。 2.5 大鼠肾脏切片免疫组织化学CD68染色 CD68为巨噬细胞的标志物，通过CD68抗体可将巨噬细胞的细胞核染色。由图4可见，正常组大鼠肾脏组织间质中少量细胞CD68阳性，实验组大鼠肾脏组织CD68阳性细胞较多，对照组大鼠肾脏组织中CD68阳性细胞最多。3组大鼠肾脏组织中CD68阳性细胞数量差异有显著性意义(P < 0.01)。"

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