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中华细胞与干细胞杂志(电子版)

中华细胞与干细胞杂志(电子版) ›› 2023, Vol. 13 ›› Issue (06) : 370 -376. doi: 10.3877/cma.j.issn.2095-1221.2023.06.007

综述

间充质干细胞体外大规模培养的研究进展
张闻宇1, 黄玉香1,†()   
  1. 1. 266000　青岛，奥克生物开发有限公司
  • 收稿日期:2023-10-17 出版日期:2023-12-01
  • 通信作者: 黄玉香

Research progress in large-scale cultivation of mesenchymal stem cells in vitro

Wenyu Zhang1, Yuxiang Huang1,†()   

  1. 1. Allcare Biomedical Development, Qingdao 266000, China
  • Received:2023-10-17 Published:2023-12-01
  • Corresponding author: Yuxiang Huang
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张闻宇, 黄玉香. 间充质干细胞体外大规模培养的研究进展[J]. 中华细胞与干细胞杂志(电子版), 2023, 13(06): 370-376.

Wenyu Zhang, Yuxiang Huang. Research progress in large-scale cultivation of mesenchymal stem cells in vitro[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2023, 13(06): 370-376.

间充质干细胞（MSCs）是一类具有自我更新和多向分化潜能的干细胞，具有免疫调节和组织修复等功能。随着MSCs在临床中的广泛应用，对细胞体外扩增以获取大量临床级细胞的需求也日益增多。因此，如何在体外获得足够数量的MSCs，是MSCs培养、制备及临床应用亟待解决的核心问题。本文总结了当前MSCs二维和三维大规模培养方法的优点、局限性以及实际应用，并对规模化、自动化和智能化培养体系的发展趋势进行展望。

关键词: 间充质干细胞, 二维培养, 三维培养, 大规模培养

Mesenchymal stem cells are a type of stem cells with self-renewal and multi- directional differentiation potential, which have functions such as immune regulation and tissue repair. With the wide application of mesenchymal stem cells in clinical practice, there is an increasing demand for cell expansion in vitro to obtain many clinical-grade cells. Therefore, getting a sufficient quantity of mesenchymal stem cells in vitro is the core problem to be solved in mesenchymal stem cell culture, preparation and clinical application. In this paper, we summarize the advantages and limitations of the current two-dimensional and three-dimensional large-scale cultivation methods for mesenchymal stem cells and the practical applications of large-scale cultivation. It also looks forward to the development trends of large-scale, automatic and intelligent cultivation systems.

Key words: Mesenchymal stem cells, Two-dimensional culture, Three-dimensional culture, Large-scale cultivation
表1 2D及3D大规模培养方法的优点与局限性
培养方式 优点 局限性
2D大规模培养 多层细胞培养瓶培养 生产工艺变动小；细胞不存在剪切损伤 放大规模有限；成本较高
  细胞工厂培养 培养环境安全；污染风险小；可实现大规模培养 层数较多的细胞工厂需要机械辅助来支持培养基的添加和移除
3D大规模培养 微载体培养 降低生产成本；污染风险小；可实现大规模培养及自动化生产； 容易聚团；细胞存在剪切损伤
  微囊化培养 防止培养过程中的聚团；细胞不存在剪切损伤 制作过程复杂；成功率低；无法实现连续化生产
  细胞微球培养 细胞存活时间长；便于储存和运输；可实现大规模培养 制备过程复杂；制备效率低；细胞存在剪切损伤
  中空纤维系统培养 适用范围广；细胞不存在剪切损伤；可实现大规模培养 无法实时观察细胞状态；细胞收获难；胰酶消化时间长
图1 MSCs 3D培养方式示意图[5]注：a图为固体球形或圆盘状颗粒微载体；b图为由纤维素或明胶制备的多孔微载体；c图为包裹凝胶的微囊体；d图为3D培养形成的球状细胞聚集体；e图为中空纤维。Cross-Sectional View为横剖面视图；Nutrients In为营养物质进入；Waste Out为代谢废物排出；Porous Fiber为多孔纤维；Intercapillary Space为毛细血管内空间；Extracapillary Space为毛细血管外空间
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