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[小鼠骨髓间充质干细胞向脂肪细胞分化过程中的微小RNA表达变化]

[miRNA expression change of differentiation of mice marrow mesenchymal stem cells into adipocytes].

作者信息

Ling Hong-Yan, Wen Ge-Bo, Hu Bi, Feng Shui-Dong, Zhang Kai-Fang, Yang Si-Si, He Jian-Qin, Yin Wei-Lan, Liao Duan-Fang

机构信息

Department of Physiology, University of South China, Hengyang 421001, China.

出版信息

Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2011 Nov;27(4):391-5.

Abstract

OBJECTIVE

To explore miRNA expression change of differentiation of mice marrow mesenchymal stem cells (MSCs) into adipocytes, which lay the foundation for further studies on molecular mechanism of miRNA regulating the differentiation of MSCs into adipocytes.

METHODS

C57BL/6 mice MSCs were isolated, cultured through the whole bone marrow method, amplified by the differential adherent method. Cell growth was observed by morphology and the expression of superficial antigen CD29, CD44, CD34 were detected through immunohistochemistry. MSCs was induced to differentiation into adipocytes with adipocyte differentiation medium, and adipogenic differentiation of MSCs was analyzed by oil Red O staining. MicroRNA microarray was used to investigate the differentially expressed miRNAs in MSCs and adipocytes.

RESULTS

(1) The fifth passage of MSCs had high purity under an inverted m icroscope. Immunohistochemistry staining showed that CD29, CD44 were positive and CD34 was negative in more than 90% MSCs. There were a large number of lipid droplets in cytoplasm after MSCs were induced with adipocyte differentiation medium, Oil O staining was positive. (2) The microarray experiment showed that 75 differentially expressed miRNAs were obtained in adipocytes compared with MSCs, 20 up-regulated and 55 down-regulated miRNAs were observed among them.

CONCLUSION

There was a expression change of miRNA of differentiation of MSCs into adipocytes, some miRNAs might play important roles in MSCs adipogenic differentiation.

摘要

目的

探讨小鼠骨髓间充质干细胞(MSCs)向脂肪细胞分化过程中miRNA的表达变化,为进一步研究miRNA调控MSCs向脂肪细胞分化的分子机制奠定基础。

方法

采用全骨髓法分离培养C57BL/6小鼠MSCs,通过差异贴壁法进行扩增。通过形态学观察细胞生长情况,采用免疫组织化学法检测表面抗原CD29、CD44、CD34的表达。用脂肪细胞分化培养基诱导MSCs向脂肪细胞分化,通过油红O染色分析MSCs的成脂分化情况。利用miRNA芯片研究MSCs和脂肪细胞中差异表达的miRNA。

结果

(1)倒置显微镜下可见第5代MSCs纯度较高。免疫组织化学染色显示,超过90%的MSCs中CD29、CD44呈阳性,CD34呈阴性。用脂肪细胞分化培养基诱导MSCs后,细胞质内出现大量脂滴,油红O染色呈阳性。(2)芯片实验显示,与MSCs相比,脂肪细胞中获得了75个差异表达的miRNA,其中20个上调,55个下调。

结论

MSCs向脂肪细胞分化过程中存在miRNA表达变化,部分miRNA可能在MSCs成脂分化中发挥重要作用。

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