Transplantation of islet-like cells from human umbilical cord mesenchymal stem cells at different induced stages for the treatment of diabetic mellitus

doi:10.3969/j.issn.2095-4344.2017.29.019

Abstract

Abstract:

BACKGROUND: In previous studies, the human umbilical cord mesenchymal stem cells (hUC-MSCs) have been successfully differentiated into islet-like cells in vitro, and insulin expressions have been found.
OBJECTIVE: To compare the therapeutic effects of different induced stages of islet-like cells differentiated from hUC-MSCs in a diabetic rat in vivo, so as to find the most suitable induced time in vitro and provide a theoretical basis for clinical treatment of diabetes mellitus.
METHODS: Passage 3 hUC-MSCs were differentiated into islet-like cells after 7, 14, 21, 28 days of oriented induction. Eighty male healthy Wistar rats, clean grade, were used in the study. Except eight rats in normal control group, all the rats were injected with streptozotocin at a dose of 70 mg/kg to establish diabetic models. The rats at 10 days after successful modeling were randomly divided into model control group, non-induced group, 7-day induction group, 14-day induction group, 21-day induction group and 28-day induction group. Rats in the normal control group and model control group were given 2 mL of culture medium without any cells and rats in the other groups were implanted with corresponding cell suspension (2×10⁶ cells) via tail vein for two sessions with an interval of 2 weeks. The blood glucose level, body mass and serum insulin level were detected during the treatment process. The rats were executed to observe the structure changes of each organ at 4 weeks after the second cell transplantation.
RESULTS AND CONCLUSION: (1) Compared with the model control group, the body mass and the serum insulin level significantly increased and the blood glucose levels significantly decreased in all the transplantation groups (P < 0.05), and the therapeutic effect was best in the 28-day induction group. (2) Compared with the model control group and normal control group, the frozen sections in all the transplantation groups showed that the morphological structures of the liver and kidney were clear with no abnormal changes, such as necrosis and fibrosis, after transplantation. These experimental results show that it is relatively safe and effective to transplant the different induced stages of islet-like cells induced by hUC-MSCs in the treatment of diabetes mellitus, and the therapeutic effect of islet-like cells at 28 days of in vitro induction is most obvious.

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: Diabetes Mellitus, Umbilical Cord, Mesenchymal Stem Cells, Cell Differentiation, Insulin-Secreting Cells, Tissue Engineering

CLC Number:

R394.2

Shan Xia, Cui Xiao-lan, Shi Han, Shen Yi, Wang Jia, Zhang She-yi, Guo Bo, Wang Yi-zhong. Transplantation of islet-like cells from human umbilical cord mesenchymal stem cells at different induced stages for the treatment of diabetic mellitus[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(29): 4703-4708.

Figures/Tables 1

2.1 实验动物数量分析除模型对照组在后期死亡3只大鼠，其他各组大鼠的生命体征较好，无脱失，均进入结果分析。 2.2 一般状态所有大鼠移植前后生命体征平稳。正常对照组大鼠精神状态良好，动作自如，反应灵敏，毛发平伏有光泽。而模型对照组大鼠精神萎靡，反应迟钝，毛发无光泽，动作迟缓，弓背蜷体，多饮多食多尿等症状明显。各治疗组大鼠精神状态、毛发光泽度较模型对照组好，多饮多食多尿等症状有所缓解。 2.3 大鼠体质量变化细胞移植治疗前，正常对照组大鼠体质量明显大于模型对照组和各治疗组，差异有非常显著性意义(P < 0.01)，模型对照组和各治疗组大鼠体质量差异均无显著性意义(P > 0.05)。细胞移植治疗后，随治疗时间延长，模型对照组糖尿病大鼠体质量逐渐下降，各治疗组大鼠体质量出现不同程度升高，与同期模型对照组相比，细胞移植7 d后，诱导28 d组的大鼠体质量显著升高(P < 0.01)，其他各治疗组大鼠体质量从第14天起显著升高(P < 0.01)，但均未恢复正常，与正常对照组相比差异有非常显著性意义(P < 0.01)；治疗后第35天起，诱导28 d组大鼠体质量显著高于其他各治疗组，差异有非常显著性意义(P < 0.01)，见图1。由此可见，各治疗组均可不同程度的升高大鼠体质量水平，而诱导28 d组体质量上升比其他各组明显。 2.4 血糖水平造模前大鼠空腹血糖水平均处于正常范围(2.80-7.56 mmol/L[18])；造模后，正常对照组大鼠血糖水平处于正常范围，而模型对照组和各治疗组大鼠血糖水平大于16.7 mmol/L，与正常对照组相比差异有非常显著性意义(P < 0.01)，模型对照组与各治疗组之间大鼠血糖水平差异无显著性意义(P > 0.05)。细胞移植治疗后，与模型对照组相比，各治疗组大鼠血糖从治疗第14天起显著降低，其中诱导21 d组和诱导28 d组从治疗第7天起显著降低，差异有非常显著性意义 (P < 0.01)，但均未恢复正常，与正常对照组相比差异有非常显著性意义(P < 0.01)；治疗21 d以后，诱导28 d组大鼠血糖水平显著低于其他各治疗组，差异有非常显著性意义(P < 0.01)，见图2。由此可见，各治疗组细胞移植后均可不同程度的降低大鼠血糖，而诱导28 d组降糖效果优于其他各组。 2.5 大鼠血清胰岛素水平治疗42 d后，与正常对照组相比，模型对照组空腹及经葡萄糖刺激后血清胰岛素水平明显下降(P < 0.01)，处于低分泌状态；与模型对照组相比，各治疗组大鼠空腹及腹腔注射葡萄糖2 h后血清胰岛素水平明显升高(P < 0.01)，其中腹腔注射葡萄糖2 h后，诱导28 d组血清胰岛素水平明显高于其他各治疗组，差异有显著性意义(P < 0.05)，见图3。由此可见，细胞移植治疗均可不同程度地提高糖尿病大鼠的血清胰岛素水平，提高葡萄糖刺激的反应性，一定程度上改善胰岛功能，而且诱导28 d组胰岛素升高水平较其他各组明显。 2.6 大鼠肝肾组织病理形态人脐带间充质干细胞治疗后42 d病理切片显示，未诱导组、诱导7 d组、诱导14 d组、诱导21 d组和诱导28 d组的大鼠肝肾组织结构清晰可见，均未发生坏死，纤维化，尚未发现肿瘤组织形成。与正常对照组和模型对照组比较，移植后各组大鼠肝肾组织形态无显著性异常改变，见图4和图5。"

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