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目的探讨携带micro-dystrophin基因的自体骨髓间充质干细胞移植入mdx鼠后在移植鼠体内分化为肌细胞的可能机制。方法采用逆转录病毒介导micro-dystrophin基因转染mdx小鼠MSCs(mMSCs),通过尾静脉注射移植治疗mdx鼠,在移植后免疫荧光检测micro-dystrophin的表达并在不同时间点检测MyoD的表达。结果移植后成功检测到micro-dystrophin,其表达随着移植时间的延长而增加;随移植时间延长MyoD阳性肌纤维比例增加,分别达到9%(4周时)、15%(8周时)、28%(12周时)。RT-PCR和Westernblot也发现,随着移植时间的延长,MyoD表达增加。结论自体mMSCs可携带外源性micro-dystrophin基因在受体鼠体内分化为micro-dystrophin阳性肌细胞,移植入的干细胞向肌细胞的分化是一个持久的、连续的过程,成肌调节因子在调节其分化过程中发挥了重大作用。
Objective To investigate the possible mechanism of autologous bone marrow mesenchymal stem cells carrying micro-dystrophin gene transplanted into mdx mice and differentiate into muscle cells in transplanted mice. Methods Mdx mouse MSCs (mMSCs) were transfected with retrovirus-mediated micro-dystrophin gene and mdx mice were transplanted via tail vein. The expression of micro-dystrophin was detected by immunofluorescence and MyoD expression at different time points after transplantation . Results Micro-dystrophin was detected successfully after transplantation. The expression of micro-dystrophin increased with the time of transplantation. The percentage of MyoD positive myofibers increased with the time of transplantation, reaching 9% (4 weeks), 15% (8 weeks), 28 % (At 12 weeks). RT-PCR and Western blotting also showed that MyoD expression increased with the extension of transplantation time. Conclusion The autologous mMSCs can differentiate into micro-dystrophin positive myogenic cells in vitro. The differentiation of stem cells into myocytes is a long-lasting and continuous process. The regulation of myogenic regulatory factors Its differentiation has played a significant role.