Stirred Suspension Culture Improves Embryoid Body Formation and Cardiogenic Differentiation of Genetically Modified Embryonic Stem Cells

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  • He Wenjun
    Department of Cell Engineering, Institute of Biotechnology, Academy of Military Medical Sciences Wound Healing and Cell Biology Laboratory, The First Affiliated Hospital, Chinese PLA General Hospital
  • Ye Lingling
    Department of Cell Engineering, Institute of Biotechnology, Academy of Military Medical Sciences
  • Li Shichong
    Department of Cell Engineering, Institute of Biotechnology, Academy of Military Medical Sciences
  • Liu Hong
    Department of Cell Engineering, Institute of Biotechnology, Academy of Military Medical Sciences
  • Wang Qiwei
    Department of Cell Engineering, Institute of Biotechnology, Academy of Military Medical Sciences
  • Fu Xiaobing
    Wound Healing and Cell Biology Laboratory, The First Affiliated Hospital, Chinese PLA General Hospital Institute of Basic Medical Sciences, Chinese PLA General Hospital
  • Han Weidong
    Institute of Basic Medical Sciences, Chinese PLA General Hospital
  • Chen Zhaolie
    Department of Cell Engineering, Institute of Biotechnology, Academy of Military Medical Sciences

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説明

Embryonic stem cells (ESCs) can propagate unlimitedly in vitro and differentiate into cardiomyocytes, which have been proposed as unlimited cell sources for cardiac cell therapy. This was limited by difficulties in large-scale generation of pure cardiomyocytes. In this study, we used stirred bioreactors to optimize the differentiation condition for mass production of embryoid bodies (EBs) derived from genetically modified mouse ESCs. Stirred suspension culture could more efficiently produce EBs and have a more uniform EB population without large necrotic centers, compared with the conventional static culture. Importantly, the cardiac-specific gene expressions (GATA binding protein 4, α-cardiac myosin heavy chain and myosin light chain-2v) were increased within EBs cultured in stirred bioreactor. Stirred suspension culture significantly increased the proportion of spontaneously contracting EBs, yielded a greater percentage of α-sarcomeric actinin-positive cells detected via flow cytometry, and harvested relatively more cardiomyocytes after G418 selection. Stirred suspension culture provided a more ideal culture condition facilitating the growth of EBs and enhancing the cardiogenic differentiation of genetically modified ESCs, which may be valuable in large-scale generation of pure cardiomyocytes.

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