Modulation of Wnt and Activin/Nodal supports efficient derivation, cloning and suspension expansion of human pluripotent stem cells

Zongyong Ai; Baohua Niu; Kui Duan; Chenyang Si; Sile Wang; Lifeng Xiang; Xiaoqing Zhu; Qingyuan Zhu; Chun Feng; Yu Yin; Shumei Zhao; Ruize Kong; Weizhi Ji; Tianqing Li

doi:10.1016/j.biomaterials.2020.120015

Modulation of Wnt and Activin/Nodal supports efficient derivation, cloning and suspension expansion of human pluripotent stem cells

Biomaterials. 2020 Aug:249:120015. doi: 10.1016/j.biomaterials.2020.120015. Epub 2020 Apr 8.

Authors

Zongyong Ai¹, Baohua Niu², Kui Duan³, Chenyang Si³, Sile Wang³, Lifeng Xiang⁴, Xiaoqing Zhu¹, Qingyuan Zhu², Chun Feng¹, Yu Yin³, Shumei Zhao³, Ruize Kong⁵, Weizhi Ji³, Tianqing Li⁶

Affiliations

¹ Yunnan Key Laboratory of Primate Biomedical Research, Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming, Yunnan, 650500, China; Yunnan Provincial Academy of Science and Technology, Kunming, Yunnan, 650500, China; XiAn ChaoYue Stem Cell Co., Ltd, No. 5 Tangxing Road, Xi'An, ShanXi, 710000, China.
² Yunnan Key Laboratory of Primate Biomedical Research, Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming, Yunnan, 650500, China.
³ Yunnan Key Laboratory of Primate Biomedical Research, Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming, Yunnan, 650500, China; Yunnan Provincial Academy of Science and Technology, Kunming, Yunnan, 650500, China.
⁴ Yunnan Key Laboratory of Primate Biomedical Research, Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming, Yunnan, 650500, China; The First People's Hospital of Yunnan Province, No.157, Jinbi Road, Xishan District, Kunming, Yunnan, 650034, China.
⁵ The First People's Hospital of Yunnan Province, No.157, Jinbi Road, Xishan District, Kunming, Yunnan, 650034, China.
⁶ Yunnan Key Laboratory of Primate Biomedical Research, Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming, Yunnan, 650500, China; Yunnan Provincial Academy of Science and Technology, Kunming, Yunnan, 650500, China; XiAn ChaoYue Stem Cell Co., Ltd, No. 5 Tangxing Road, Xi'An, ShanXi, 710000, China. Electronic address: litq@lpbr.cn.

PMID: 32311594
DOI: 10.1016/j.biomaterials.2020.120015

Abstract

Various culture systems have been used to derive and maintain human pluripotent stem cells (hPSCs), but they are inefficient in sustaining cloning and suspension expansion of hPSCs. Through systematically modulating Wnt and Activin/Nodal signaling, we developed a defined medium (termed AIC), which enables efficient cloning and long-term expansion of hPSCs (AIC-hPSCs) through single-cell passage on feeders, matrix or in suspension (25-fold expansion in 4 days) and maintains genomic stability of hPSCs over extensive expansion. Moreover, the AIC medium supports efficient derivation of hPSCs from blastocysts or somatic cells under feeder-free conditions. Compared to conventional hPSCs, AIC-hPSCs have similar gene expression profiles but down-regulated differentiation genes and display higher metabolic activity. Additionally, the AIC medium shows a good compatibility for different hPSC lines under various culture conditions. Our study provides a robust culture system for derivation, cloning and suspension expansion of high-quality hPSCs that benefits GMP production and processing of therapeutic hPSC products.

Keywords: Genomic stability; Human pluripotent stem cells; Modulation of wnt and activin/nodal; Single-cell passage; Suspension expansion; β-catenin and Smad2/3.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Activins*
Cell Culture Techniques
Cell Differentiation
Cloning, Molecular
Humans
Pluripotent Stem Cells*

Substances

Activins