title: Generation of cardiac spheres from primate pluripotent stem cells in a small molecule-based 3D system.
abstract: Pluripotent stem cell (PSC) usage in heart regenerative medicine requires producing enriched cardiomyocytes (CMs) with mature phenotypes in a defined medium. However, current methods are typically performed in 2D environments that produce immature CMs. Here we report a simple, growth factor-free 3D culture system to rapidly and efficiently generate 85.07  ±  1.8% of spontaneously contractile cardiac spheres (scCDSs) using 3D-cultured human and monkey PSC-spheres. Along with small molecule-based 3D induction, this protocol produces CDSs of up to 95.7% CMs at a yield of up to 237 CMs for every input pluripotent cell, is effective for human and monkey PSCs, and maintains 81.03  ±  12.43% of CDSs in spontaneous contractibility for over three months. These CDSs displayed CM ultrastructure, calcium transient, appropriate pharmacological responses and CM gene expression profiles specific for maturity. Furthermore, 3D-derived CMs displayed more mature phenotypes than those from a parallel 2D-culture. The system is compatible to large-scaly produce CMs for disease study, cell therapy and pharmaceutics screening.
authors.affiliation: Yunnan Key Laboratory of Primate Biomedical Research; Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming, Yunnan 650500, China; National Engineering Research Center of Biomedicine and Animal Science, Kunming, Yunnan 650500, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Jiang
authors.affiliation: Yunnan Key Laboratory of Primate Biomedical Research; Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming, Yunnan 650500, China; National Engineering Research Center of Biomedicine and Animal Science, Kunming, Yunnan 650500, China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Xiang
authors.affiliation: Yunnan Key Laboratory of Primate Biomedical Research; Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming, Yunnan 650500, China; National Engineering Research Center of Biomedicine and Animal Science, Kunming, Yunnan 650500, China.
authors.forename: Zongyong
authors.initials: Z
authors.lastname: Ai
authors.affiliation: Yunnan Key Laboratory of Primate Biomedical Research; Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming, Yunnan 650500, China; National Engineering Research Center of Biomedicine and Animal Science, Kunming, Yunnan 650500, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.affiliation: Yunnan Key Laboratory of Primate Biomedical Research; Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming, Yunnan 650500, China; National Engineering Research Center of Biomedicine and Animal Science, Kunming, Yunnan 650500, China.
authors.forename: Yanjiao
authors.initials: Y
authors.lastname: Li
authors.affiliation: Yunnan Key Laboratory of Primate Biomedical Research; Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming, Yunnan 650500, China; National Engineering Research Center of Biomedicine and Animal Science, Kunming, Yunnan 650500, China. Electronic address: wji@kbimed.com.
authors.forename: Weizhi
authors.initials: W
authors.lastname: Ji
authors.affiliation: Yunnan Key Laboratory of Primate Biomedical Research; Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming, Yunnan 650500, China; National Engineering Research Center of Biomedicine and Animal Science, Kunming, Yunnan 650500, China. Electronic address: litq@kbimed.com.
authors.forename: Tianqing
authors.initials: T
authors.lastname: Li
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology*; D006801:Humans; D008252:Macaca fascicularis; D009200:Myocardial Contraction; D032383:Myocytes, Cardiac / Q000166:cytology*; D039904:Pluripotent Stem Cells / Q000166:cytology*; D018874:Spheroids, Cellular / Q000166:cytology
keywords: 3D suspension culture of plurioptent stem cells; Cardiac maturity; Cardiac spheres; Primate embryonic stem cells; Small molecule-based 3D differentiation