体外两种方式培养的肠神经干细胞生物学特性比较

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Abstract Objective: To find a suitable proliferation way of culturing enteric neural stem cells in vitro, and to compare the biological characteristics of stem cells which were cultured in dishes coated and uncoated with ﬁbronectin (FN). Methods: Enteric neural stem cells were isloated from gut of 15 days rat embryos, the neurospheres of fifth days were extracted from two groups. Growth status, proliferation and differentiation capacity of cells were compared through morphological observation, immunofluorescence cytochemistry and flow cytometer analysis between the two groups. Results: Compared with the FN coated group, the neurospheres of fifth days were the state of semi-adherent growth, the neurospheral morphology was regular, and were beginning to get dark in the center of them, and the spheres did not form a large amount of nerve fiber. Under fluorescence microscope, the typical neurospheres which were shown positive to BrdU in two groups were found. Radial nerve fiber of the neurospheres were positive to Tuj-1 (neuronal axons). After culturing for 5 days, compared with treatment with FN group (12.67±2.16)%, flow cytometer analysis showed positive to Nestin of the FN uncoated group had inacresed significantly (20.69±2.15)% (P<0.05); BrdU which mark proliferative cell increased significantly (35.82±2.11)% vs FN group (12.69±1.13)% (P<0.05); The percentage of Tuj-1 positive to neuron decreased significantly (10.67±1.14)% vs FN group (21.77±2.53)% (P<0.05). Conclusion: The way of uncoating culture reduces the time of forming neurospheres which were regular shape; maintains stem cell self-proliferation, reduce enteric neural stem cell differentiation and is suitable for stem cell self-proliferation in vitro.

Key words： enteric neural stem cells proliferation culture fibronectin

Received: 16 September 2014

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	Articles by authors
	LIU Miaoqing
	WANG Yongbiao
	HUANG Xiaozhong
	ZHU Libin
	CHEN Xiaoming
	LI Zhongrong.

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https://xb.wmu.edu.cn/EN/ OR https://xb.wmu.edu.cn/EN/Y2015/V45/I8/566

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