人外周血单个核细胞体外重编程为少突胶质前体细胞研究

doi:10.3969/j.issn.1006-7795.2017.04.013

首都医科大学学报 ›› 2017, Vol. 38 ›› Issue (4): 553-558.doi: 10.3969/j.issn.1006-7795.2017.04.013

人外周血单个核细胞体外重编程为少突胶质前体细胞研究

唐玺和^1,2,3,4, 李默^2,3, 王淑艳^2,3, 李鹏燕^2,3, 张愚^2,3, 陈志国^2,3, 陈惠^1,3,4

1. 中国康复医学研究所, 北京 100053;
2. 首都医科大学宣武医院细胞治疗室, 北京 100053;
3. 北京脑重大疾病研究院神经损伤和修复所, 北京 100053;
4. 北京神经损伤和康复重点实验室, 北京 100053

收稿日期:2016-06-14 出版日期:2017-07-21 发布日期:2017-07-20
通讯作者: 陈志国,陈惠 E-mail:chenzhiguo@gmail.com;chenhui55299@163.com
基金资助:
国家自然科学基金（81661130160），北京市科委计划项目（Z151100001615055）

Generation of oligodendrocyte progenitor cells from human peripheral blood mononuclear cells

Tang Xihe^1,2,3,4, Li Mo^2,3, Wang Shuyan^2,3, Li Pengyan^2,3, Zhang Y. Alex^2,3, Chen Zhiguo^2,3, Chen Hui^1,3,4

1. China Rehabilitation Research Center, Beijing 100053, China;
2. Cell Therapy Department, Xuanwu Hospital, Capital Medical University, Beijing 100053, China;
3. Center of Neural Injury and Repair, Beijing Institute for Brain Disorders, Beijing 100053, China;
4. Beijing Key Laboratory of Neural Injury and Rehabilitation, Beijing 100053, China

Received:2016-06-14 Online:2017-07-21 Published:2017-07-20
Supported by:
This study was supported by National Natural Science Foundation of China(81661130160), Beijing Municipal Science and Technology Commission (Z151100001615055)

摘要/Abstract

摘要： 目的利用非整合质粒载体在体外将成人外周血中单个核细胞重编程为少突胶质前体细胞（oligodendrocyte progenitor cells，OPCs）。方法经外周静脉采集志愿者血液5 mL，利用Ficoll-Paquem密度梯度离心法获得单个核细胞，体外扩增培养后，电转染携带外源基因（OCT4，SOX2，KLF4，C-myc，LIN28，Nanog）的质粒，然后在加有化学小分子的特定培养基中分三步培养。结果转染后30 d左右，可以获得血小板衍生生长因子受体α（platelet-derived growth factor receptor-α，PDGFR-α）阳性的早期少突胶质前体细胞（Pre-OPC），该细胞能传20 代以上，继续分化30 d左右可以获得表达O4的少突胶质前体细胞。结论利用非整合质粒载体携带外源基因可以将成人外周血单个核细胞在较短期时间内重编程为具有增生能力的早期少突胶质前体细胞，且能继续分化为少突胶质前体细胞。

关键词: 外周血单个核细胞, 重编程, 少突胶质前体细胞

Abstract: Objective To generate oligodendrocyte progenitor cells (OPCs) from peripheral blood mononuclear cells by episomal vectors.Methods Peripheral blood of a donor was collected by venipuncture and then mononuclear cells (MNCs) were isolated by density-based centrifugal separation. After a short period of expansion, the isolated MNCs were transfected with three plasmids expressing OCT4, SOX2, KLF4, C-MYC, LIN28, NANOG, and cultured in chemical defined medium with small molecules. Results Sixty days later, the O4 oligodendrocyte progenitor cells appeared and could be expanded more than 60 passages. Conclusion The peripheral blood MNCs can be converted to oligodendrocyte progenitor cells by non-integrative plasmid vectors.

Key words: peripheral blood mononuclear cells, cell reprogramming, oligodendrocyte progenitor cells

中图分类号:

R329.2

唐玺和, 李默, 王淑艳, 李鹏燕, 张愚, 陈志国, 陈惠. 人外周血单个核细胞体外重编程为少突胶质前体细胞研究[J]. 首都医科大学学报, 2017, 38(4): 553-558.

Tang Xihe, Li Mo, Wang Shuyan, Li Pengyan, Zhang Y. Alex, Chen Zhiguo, Chen Hui. Generation of oligodendrocyte progenitor cells from human peripheral blood mononuclear cells[J]. Journal of Capital Medical University, 2017, 38(4): 553-558.

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人外周血单个核细胞体外重编程为少突胶质前体细胞研究

Generation of oligodendrocyte progenitor cells from human peripheral blood mononuclear cells

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