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[1]顾敬超,王明连,王群,等.一株用于研究环境因素暴露效应的神经样细胞[J].应用与环境生物学报,2017,23(03):482-485.[doi:2016.08006]
 GU Jingchao,WANG Minglian,WANG Qun,et al.Immortal neural-like cells to study the biological effects of exposure to environmental factors[J].Chinese Journal of Applied & Environmental Biology,2017,23(03):482-485.[doi:2016.08006]
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一株用于研究环境因素暴露效应的神经样细胞()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
23卷
期数:
2017年03期
页码:
482-485
栏目:
研究论文
出版日期:
2017-06-25

文章信息/Info

Title:
Immortal neural-like cells to study the biological effects of exposure to environmental factors
作者:
顾敬超王明连王群李艳刘强强路通曾毅
1北京工业大学生命科学与生物工程学院 北京 100124 2北京工业大学材料科学与工程学院 北京 100124 3中国疾病预防控制中心病毒病预防控制所 北京 100052
Author(s):
GU Jingchao WANG Minglian WANG Qun LI Yan LIU Qiangqiang LU Tong & ZENG Yi
1College of Life Science and Bioengineering, Beijing University of Technology, Beijing, 100124, China 2College of Material Science and Engineering, Beijing University of Technology, Beijing, 100124, China 3National Institute for Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, 100052, China
关键词:
间充质干细胞诱导分化神经样细胞永生化环境暴露
Keywords:
mesenchymal stem cells induced differentiation neural-like cell immortalized environmental exposure
分类号:
X171.5
DOI:
2016.08006
摘要:
为建立一株可用于研究长期环境因素暴露的永生化神经样细胞,使用bFGF作为预诱导剂,BHA、DMSO和β-巯基乙醇作为诱导剂,诱导人脐带间充质干细胞向神经样细胞分化,观察细胞形态变化,并通过细胞免疫荧光和RT-PCR检测Nestin、NSE、GFAP和MAP2的表达,判断是否具有神经样细胞特异性. 将诱导成功的细胞连续传代,利用稀释法分离出单细胞克隆. 筛选出可以连续传代50次以上的细胞,RT-PCR检测Nestin和MAP2基因表达差异,判断所获得的单克隆细胞的神经样特性是否能够维持. 结果显示,诱导后的细胞胞体凸起,免疫荧光检测Nestin、NSE、GFAP蛋白表达均阳性,说明成功将MSCs诱导为神经样细胞,Nestin基因表达在传代11次时升高1.32倍,在50次时降低8.85倍,而MAP2基因表达在传代11次时升高2.12倍,在50次时升高7.29倍,说明其已经由神经干细胞分化为神经样细胞. 本研究得到的神经样细胞单克隆可以连续传代,并且保持神经样细胞特性,可以作为检测环境因素对神经系统慢性损伤作用研究的细胞模型. (图5 表1 参21)
Abstract:
In this study, we aimed to establish an immortal neural-like cell strain that could be used to study the effects of long-term exposure to environmental factors. After preconditioning with bFGF, human umbilical cord-derived mesenchymal stem cells (MSCs) were then induced with BHA, DMSO, and β-mercaptoethanol. Changes in cell morphology were observed under microscope and the expression levels of Nestin, NSE, GFAP, and MAP2 were determined by immunofluorescence and real-time PCR to estimate nerve cell specificity. The successfully induced cells were serially passaged, and single-cell clones were isolated. A cell clone that was subcultured more than 50 times was selected. Based on Nestin and MAP2 expression levels, we obtained a cell line with nerve-like properties. In addition, the induced cells showed surface projections, and the expression of Nestin, NSE, and GFAP indicated that the MSCs had differentiated into neuron-like cells. Nestin expression increased 1.32-fold at the 11th subculture and decreased 8.85-fold at the 50th subculture. Specifically, MAP2 expression increased 2.12- and 7.29-fold at the 11th and 50th subculture, respectively, confirming that the neural stem cells differentiated into neuron-like cells that exhibited the neuron-like cell properties. The monoclonal neural-like cells could be serially passaged, and they maintained their neuron-like properties. Thus, these cells can be used to study the chronic effects of long-term environmental exposures on the nervous system.

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更新日期/Last Update: 2017-06-25