神经营养因子-3联合全反式维甲酸与β-巯基乙醇诱导大鼠骨髓间充质干细胞分化效果比较

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Abstract Objective: To explore the differences between two methods of inducing BMSCs into neuron-like cells in vitro. Methods: BMSCs were cultured and sub-cultured by the main method of whole bone marrow directly adherent to flasks, and the surface markers CD29, CD90 and CD45 of the third generation were then detected by flow cytometry. The third generation of BMSCs were pre-induced by basic fibroblast growth factor(bFGF), then neurotrophin-3 (NT-3)+retinoic acid (RA) and beta-mercaptoethanol (β-BME) were added respectively. Immunofluorescence was used to identify whether cells expressed neuron-specific enolase (NSE). Results: BMSCs with high quantity and purity could be obtained by the method of whole bone marrow directly adherent to flasks. We observed that the cells expressed CD29 and CD90 but not CD45 through flow cytometric detection, confirming that the cells were BMSCs. Both groups expressed NSE. The positive rate of 7 d in (NT-3+RA) group was higher than other time points (P<0.01), and the positive rate of 5 h in the β-BME group was higher than 1 h, and the positive rate of 1 h and 5 h in (NT-3+RA) group was lower than the β-BME group (P<0.01), but almost all cells in the β-BME group died in 24 h. Conclusion: The method of whole bone marrow directly adherent to flasks is suitable for extracting BMSCs. Both methods can be used to induce BMSCs differentiation into neuron-like cells. The survival time and later-stage positive rate of (NT-3+RA) group is better than β-BME group.

Key words： bone marrow mesenchymal stem cells neuron-like cells differentiation rats

Received: 15 January 2018

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	ZHANG Jie
	QIN Huali
	JIANG Ming.

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https://xb.wmu.edu.cn/EN/ OR https://xb.wmu.edu.cn/EN/Y2018/V48/I9/641

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