title: Stirred suspension culture improves embryoid body formation and cardiogenic differentiation of genetically modified embryonic stem cells.
abstract: 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.
authors.affiliation: Department of Cell Engineering, Institute of Biotechnology, Academy of Military Medical Sciences, Beijing, China.
authors.forename: Wenjun
authors.initials: W
authors.lastname: He
authors.forename: Lingling
authors.initials: L
authors.lastname: Ye
authors.forename: Shichong
authors.initials: S
authors.lastname: Li
authors.forename: Hong
authors.initials: H
authors.lastname: Liu
authors.forename: Qiwei
authors.initials: Q
authors.lastname: Wang
authors.forename: Xiaobing
authors.initials: X
authors.lastname: Fu
authors.forename: Weidong
authors.initials: W
authors.lastname: Han
authors.forename: Zhaolie
authors.initials: Z
authors.lastname: Chen
mesh_terms: D000818:Animals; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002460:Cell Line; D058732:Embryoid Bodies / Q000166:cytology*; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology*; D013535:Suspensions
chemical_list: D013535:Suspensions