A scalable culture system incorporating microcarrier for specialised mesenchymal stem cells from human embryonic stem cells

Tingting Gao; Xiyuan Zhao; Jie Hao; Yao Tian; Huike Ma; Wenjing Liu; Bin An; Faguo Sun; Shasha Liu; Baojie Guo; Shuaishuai Niu; Zhongwen Li; Chenxin Wang; Yukai Wang; Guihai Feng; Liu Wang; Wei Li; Jun Wu; Meijin Guo; Qi Zhou; Qi Gu

doi:10.1016/j.mtbio.2023.100662

A scalable culture system incorporating microcarrier for specialised mesenchymal stem cells from human embryonic stem cells

Mater Today Bio. 2023 May 5:20:100662. doi: 10.1016/j.mtbio.2023.100662. eCollection 2023 Jun.

Authors

Tingting Gao^{1

2}, Xiyuan Zhao^{1

3

2}, Jie Hao^{4

3

2}, Yao Tian^{1

2}, Huike Ma⁵, Wenjing Liu³, Bin An¹, Faguo Sun^{1

2}, Shasha Liu^{1

2}, Baojie Guo^{1

2}, Shuaishuai Niu^{4

3}, Zhongwen Li^{1

3}, Chenxin Wang^{1

3}, Yukai Wang^{1

3

2}, Guihai Feng^{1

3

2}, Liu Wang^{1

3

2}, Wei Li^{1

3

2}, Jun Wu^{1

3}, Meijin Guo⁵, Qi Zhou^{1

3

2}, Qi Gu^{1

3

2}

Affiliations

¹ State Key Laboratory of Stem Cell and Reproductive Biology, State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
² University of Chinese Academy of Sciences, Beijing 100049, China.
³ Beijing Institute for Stem Cell and Regenerative Medicine, Beijing 100101, China.
⁴ National Stem Cell Resource Center, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
⁵ State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.

Abstract

Mesenchymal stromal cells (MSCs) derived from human embryonic stem cells (hESCs) are a desirable cell source for cell therapy owing to their capacity to be produced stably and homogeneously in large quantities. However, a scalable culture system for hPSC-derived MSCs is urgently needed to meet the cell quantity and quality requirements of practical clinical applications. In this study, we developed a new microcarrier with hyaluronic acid (HA) as the core material, which allowed scalable serum-free suspension culture of hESC-derived MSCs (IMRCs). We used optimal microcarriers with a coating collagen concentration of 100 μg/mL or concave-structured surface (cHAMCs) for IMRC amplification in a stirred bioreactor, expanding IMRCs within six days with the highest yield of over one million cells per milliliter. In addition, the harvested cells exhibited high viability, immunomodulatory and regenerative therapeutic promise comparable to monolayer cultured MSCs while showing more increased secretion of extracellular matrix (ECM), particularly collagen-related proteins. In summary, we have established a scalable culture system for hESC-MSCs, providing novel approaches for future cell therapies.

Keywords: Bioreactor; Culture system; Human embryonic stem cells; Mesenchymal stem cells; Microcarrier.