Feeder-free generation and transcriptome characterization of functional mesenchymal stromal cells from human pluripotent stem cells

Lidan Luo; Yan Zhou; Chenxi Zhang; Jinrong Huang; Jie Du; Jinqi Liao; Natasja Leth Bergholt; Cody Bünger; Fengping Xu; Lin Lin; Guangdong Tong; Guangqian Zhou; Yonglun Luo

Stem Cell Research (Oct 2020)

Feeder-free generation and transcriptome characterization of functional mesenchymal stromal cells from human pluripotent stem cells

Lidan Luo,
Yan Zhou,
Chenxi Zhang,
Jinrong Huang,
Jie Du,
Jinqi Liao,
Natasja Leth Bergholt,
Cody Bünger,
Fengping Xu,
Lin Lin,
Guangdong Tong,
Guangqian Zhou,
Yonglun Luo

Affiliations

Lidan Luo: Department of Liver Disease, Shenzhen Traditional Chinese Medicine Hospital, Guangzhou University of Chinese Medicine, Shenzhen 518033, China; Department of Biomedicine, Aarhus University, 8000 Aarhus C, Denmark
Yan Zhou: Department of Biomedicine, Aarhus University, 8000 Aarhus C, Denmark; Department of Medical Cell Biology and Genetics, Guangdong Key Laboratory of Genomic Stability and Disease Prevention, Shenzhen Key Laboratory of Anti-aging and Regenerative Medicine, and Shenzhen Engineering Laboratory of Regenerative Technologies for Orthopaedic Diseases, Health Sciences Center, Shenzhen University, Shenzhen 518060, China; Lungene Technologies Co., Ltd, Shenzhen, China; Corresponding authors at: Department of Biomedicine, Aarhus University, 8000 Aarhus C, Denmark (Y. Zhou) and (Y. Luo); Department of Medical Cell Biology and Genetics, Health Sciences Center, Shenzhen University (G. Zhou).
Chenxi Zhang: Lars Bolund Institute of Regenerative Medicine, BGI-Qingdao, BGI-Shenzhen, Shenzhen 518083, China
Jinrong Huang: Lars Bolund Institute of Regenerative Medicine, BGI-Qingdao, BGI-Shenzhen, Shenzhen 518083, China; Department of Biology, University of Copenhagen, 2100 Copenhagen, Denmark
Jie Du: Department of Medical Cell Biology and Genetics, Guangdong Key Laboratory of Genomic Stability and Disease Prevention, Shenzhen Key Laboratory of Anti-aging and Regenerative Medicine, and Shenzhen Engineering Laboratory of Regenerative Technologies for Orthopaedic Diseases, Health Sciences Center, Shenzhen University, Shenzhen 518060, China; Lungene Technologies Co., Ltd, Shenzhen, China
Jinqi Liao: Department of Medical Cell Biology and Genetics, Guangdong Key Laboratory of Genomic Stability and Disease Prevention, Shenzhen Key Laboratory of Anti-aging and Regenerative Medicine, and Shenzhen Engineering Laboratory of Regenerative Technologies for Orthopaedic Diseases, Health Sciences Center, Shenzhen University, Shenzhen 518060, China; Lungene Technologies Co., Ltd, Shenzhen, China
Natasja Leth Bergholt: Department of Clinical Medicine, Aarhus University Hospital, Aarhus, Denmark
Cody Bünger: Department of Clinical Medicine, Aarhus University Hospital, Aarhus, Denmark
Fengping Xu: Lars Bolund Institute of Regenerative Medicine, BGI-Qingdao, BGI-Shenzhen, Shenzhen 518083, China
Lin Lin: Department of Biomedicine, Aarhus University, 8000 Aarhus C, Denmark
Guangdong Tong: Department of Liver Disease, Shenzhen Traditional Chinese Medicine Hospital, Guangzhou University of Chinese Medicine, Shenzhen 518033, China
Guangqian Zhou: Department of Medical Cell Biology and Genetics, Guangdong Key Laboratory of Genomic Stability and Disease Prevention, Shenzhen Key Laboratory of Anti-aging and Regenerative Medicine, and Shenzhen Engineering Laboratory of Regenerative Technologies for Orthopaedic Diseases, Health Sciences Center, Shenzhen University, Shenzhen 518060, China; Corresponding authors at: Department of Biomedicine, Aarhus University, 8000 Aarhus C, Denmark (Y. Zhou) and (Y. Luo); Department of Medical Cell Biology and Genetics, Health Sciences Center, Shenzhen University (G. Zhou).
Yonglun Luo: Lars Bolund Institute of Regenerative Medicine, BGI-Qingdao, BGI-Shenzhen, Shenzhen 518083, China; Department of Biomedicine, Aarhus University, 8000 Aarhus C, Denmark; Corresponding authors at: Department of Biomedicine, Aarhus University, 8000 Aarhus C, Denmark (Y. Zhou) and (Y. Luo); Department of Medical Cell Biology and Genetics, Health Sciences Center, Shenzhen University (G. Zhou).

Journal volume & issue: Vol. 48
p. 101990

Abstract

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Induced mesenchymal stromal cells (iMSCs) derived from human pluripotent stem cells (PSCs) are attractive cells for regenerative medicine. However, the transcriptome of iMSCs and signature genes that can distinguish MSCs from fibroblasts and other cell types are rarely explored. In this study, we reported an optimized feeder-free method for the generation of iMSCs from human pluripotent stem cells. These iMSCs display a typical MSC morphology, express classic MSC markers (CD29, CD44, CD73, CD90, CD105, CD166), are negative for lymphocyte markers (CD11b, CD14, CD31, CD34, CD45, HLA-DR), and are potent for osteogenic and chondrogenic differentiation. Using genome-wide transcriptome profiling, we created an easily accessible transcriptome reference for the process of differentiating PSCs into iMSCs. The iMSC transcriptome reference revealed clear patterns in the silencing of pluripotency genes, activation of lineage commitment genes, and activation of mesenchymal genes during iMSC generation. All previously known positive and negative markers for MSCs were confirmed by our iMSC transcriptomic reference, and most importantly, gene classification and time course analysis identified 52 genes including FN1, TGFB1, TAGLN and SERPINE1, which showed significantly higher expression in MSCs (over 3 folds) than fibroblasts and other cell types. Taken together, these results provide a useful method and important resources for developing and understanding iMSCs in regenerative medicine.

Published in Stem Cell Research

ISSN: 1873-5061 (Print); 1876-7753 (Online)
Publisher: Elsevier
Country of publisher: Netherlands
LCC subjects: Science: Biology (General)
Website: https://www.journals.elsevier.com/stem-cell-research

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