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[人骨髓间充质干细胞骨骼肌分化中肌源性标志物的表达]

[Expressions of myogenic markers in skeletal muscle differentiation of human bone marrow mesenchymal stem cells].

作者信息

Liu Tai-yun, Dai Hong, Lin Jun, Li Mei, Xiong Fu, Feng Shan-wei, Zhang Ya-ni, Zhang Cheng

机构信息

Department of Neurology, Guizhou Provincial Hospital, Guiyang 550002, China.

出版信息

Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2010 Oct;32(5):516-20. doi: 10.3881/j.issn.1000-503X.2010.05.010.

Abstract

OBJECTIVE

To investigate the expressions of myogenic markers MyoD, myogenin,and desmin in skeletal muscle differentiation of human bone marrow mesenchymal stem cells (hBM-MSCs).

METHODS

Myogenic markers MyoD, myogenin,and desmin of hBM-MSCs cultured in vitro were detected by immunofluorescence and RT-PCR. A total of 21 8-to-10 week-old immunosuppressed mdx mice were transplanted with 1x107 passage 5 of hBM-MSCs. The mice were euthanized 2-24 weeks after transplantation,and gastrocnemius muscle were analyzed for human MyoD, myogenin,desmin,and dystrophin (Dys) expressions by immunohistochemistry and RT-PCR.

RESULTS

The numbers of MyoD-,myogenin-,and desmin-positive cells per 100 hBM-MSCs were 23.5∓5.3, 30.7∓6.2, and 28.4∓5.7, respectively. MyoD, myogenin, and desmin mRNA was observed in passage 5 of hBM-MSCs. After two weeks of hBM-MSCs transplantation,a small number of MyoD-and myogenin-positive cells were observed in skeletal muscle of mdx mice,and desmin-positive cells were observed 4 weeks after transplantation. Expressions of MyoD and myogenin were detected in the muscle of mdx mice 2-4 weeks after hBM-MSCs transplantation, which reached a peak 12-16 weeks later. Desmin was expressed in the muscle of mdx mice 4-8 weeks after transplantation,with much more expression after 16 weeks of transplantation. A small number of Dys-positive cell and Dys mRNA expression were presented in the muscle of mdx mice 4 and 8 weeks after hBM-MSCs transplantation,respectively. The expression of Dys in the muscle of mdx mice increased gradually after transplantation.

CONCLUSION

hBM-MSCs have the potential of myogenic differentiation in vitro and contribute to myogenic conversion in xenogeneic animal,during which the up-regulation of MyoD and myogenin expressions may play an important role.

摘要

目的

研究人骨髓间充质干细胞(hBM-MSCs)骨骼肌分化过程中肌源性标志物MyoD、肌细胞生成素及结蛋白的表达情况。

方法

采用免疫荧光法和逆转录-聚合酶链反应(RT-PCR)检测体外培养的hBM-MSCs的肌源性标志物MyoD、肌细胞生成素及结蛋白。将1×10⁷ 第5代hBM-MSCs移植到21只8至10周龄的免疫抑制mdx小鼠体内。移植后2至24周对小鼠实施安乐死,通过免疫组织化学和RT-PCR分析腓肠肌中人类MyoD、肌细胞生成素、结蛋白及抗肌萎缩蛋白(Dys)的表达情况。

结果

每100个hBM-MSCs中MyoD、肌细胞生成素及结蛋白阳性细胞数分别为23.5±5.3、30.7±6.2和28.4±5.7。在第5代hBM-MSCs中观察到MyoD、肌细胞生成素及结蛋白的mRNA。hBM-MSCs移植两周后,在mdx小鼠骨骼肌中观察到少量MyoD和肌细胞生成素阳性细胞,移植4周后观察到结蛋白阳性细胞。hBM-MSCs移植后2至4周在mdx小鼠肌肉中检测到MyoD和肌细胞生成素的表达,12至16周后达到峰值。移植后4至8周在mdx小鼠肌肉中表达结蛋白,移植16周后表达量更多。hBM-MSCs移植后4周和8周,mdx小鼠肌肉中分别出现少量Dys阳性细胞和Dys mRNA表达。移植后mdx小鼠肌肉中Dys的表达逐渐增加。

结论

hBM-MSCs在体外具有肌源性分化潜能,并在异种动物中促成肌源性转化,在此过程中MyoD和肌细胞生成素表达上调可能起重要作用。

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