遗传 ›› 2018, Vol. 40 ›› Issue (12): 1120-1128.doi: 10.16288/j.yczz.18-123

• 技术与方法 • 上一篇    下一篇

利用微载体悬浮培养人脐带间充质干细胞

李夏1,2,3,滑慧娟2,3,郝捷2,3,王柳2,3,刘忠华1()   

  1. 1. 东北农业大学生命科学学院,哈尔滨 150030
    2. 北京干细胞库,北京 100190
    3. 中国科学院动物研究所,北京 100101
  • 收稿日期:2018-06-15 修回日期:2018-09-03 出版日期:2018-12-20 发布日期:2018-09-03
  • 通讯作者: 刘忠华 E-mail:liuzhonghua@neau.edu.cn
  • 作者简介:李夏,硕士研究生,专业方向：发育生物学。E-mail: lixia_bjscb@foxmail.com
  • 基金资助:
    中国科学院重点部署项目(编号：ZDRW-ZS-2016-2)资助

Culturing of human umbilical cord mesenchymal stem cells via a 3D suspension culture system using the microcarrier

Xia Li1,2,3,Huijuan Hua2,3,Jie Hao2,3,Liu Wang2,3,Zhonghua Liu1()   

  1. 1. College of Life Science, Northeast Agricultural University, Harbin, 150030, China
    2. Beijing Stem Cell Bank, Chinese Academy of Sciences, Beijing 100190, China
    3. Institute of Zoology, Chinese Academy of Science, Beijing 100101, China
  • Received:2018-06-15 Revised:2018-09-03 Online:2018-12-20 Published:2018-09-03
  • Contact: Liu Zhonghua E-mail:liuzhonghua@neau.edu.cn
  • Supported by:
    Supported by the Key Deployment Projects of CAS (No. ZDRW-ZS-2016-2)

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摘要：

随着干细胞研究的不断深入,干细胞功能分化研究和临床应用转化的需求日益提升。人脐带间充质干细胞(human umbilical cord mesenchymal stem cells, hUCMSCs)来源广泛,不仅自我更新能力强、能够分化成多种类型的成体细胞,而且其自身具有免疫调节能力,不易引发免疫排斥反应,在干细胞功能分化研究和临床应用中具有巨大应用前景和应用潜力。目前,传统的细胞培养方式培养效率低、细胞活性较差,不能满足日益增长的研究和应用需求。本研究利用微载体结合旋转瓶的悬浮培养方法,通过优化细胞接种量及转速等影响因素,快速获得大量高质量的人脐带间充质干细胞。经悬浮培养总细胞量可高达到7×10 8个细胞/L,而且细胞活性较高,MSC 特异性标记物表达良好,在恢复平面培养后仍能维持MSC的正常细胞形态和增殖能力。高效脐带间充质干细胞悬浮培养体系的初步建立,为未来的干细胞功能分化研究和临床应用奠定了基础。

关键词: 人脐带间充质干细胞, 3D悬浮培养体系, 平面(2D)培养体系, 微载体

Abstract:

With the rapid development of stem cell research, the demands for high quality stem cells in cell differentiation studies, cell therapy and clinical applications increase significantly. Human umbilical cord mesenchymal stem cells (hUCMSCs) hold great potentials in these applications due to its wide availability, self-renewal capacity, good pluripotency, and in particular, self-immunomodulation ability. However, the conventional adherent culture system remains challenging in mass production of high-quality stem cells. In this study, we set up a 3D suspension culture system by using spinner flasks added with microcarrier. By optimizing the seeding density and rotation speed, we achieved a cell density as higher as 7×10 8cells/L per vessel. The cultured MSCs had good viability, and the expression levels of the MSC markers were similar to those of 2D-cultured MSCs. After being transferred back into a 2D culture system, the MSCs still maintained normal morphology and proliferation ability. These results indicated that the 3D suspension culture system established in this study provides a fundamental basis for mass production of high-quality MSCs for future research and clinical applications.

Key words: human umbilical cord mesenchymal stem cells (hUCMSCs), 3D Suspension culture system, Adherent (2D) culture system, Microcarrier