title: Induction and enhancement of cardiac cell differentiation from mouse and human induced pluripotent stem cells with cyclosporin-A.
abstract: Induced pluripotent stem cells (iPSCs) are novel stem cells derived from adult mouse and human tissues by reprogramming. Elucidation of mechanisms and exploration of efficient methods for their differentiation to functional cardiomyocytes are essential for developing cardiac cell models and future regenerative therapies. We previously established a novel mouse embryonic stem cell (ESC) and iPSC differentiation system in which cardiovascular cells can be systematically induced from Flk1(+) common progenitor cells, and identified highly cardiogenic progenitors as Flk1(+)/CXCR4(+)/VE-cadherin(-) (FCV) cells. We have also reported that cyclosporin-A (CSA) drastically increases FCV progenitor and cardiomyocyte induction from mouse ESCs. Here, we combined these technologies and extended them to mouse and human iPSCs. Co-culture of purified mouse iPSC-derived Flk1(+) cells with OP9 stroma cells induced cardiomyocyte differentiation whilst addition of CSA to Flk1(+) cells dramatically increased both cardiomyocyte and FCV progenitor cell differentiation. Spontaneously beating colonies were obtained from human iPSCs by co-culture with END-2 visceral endoderm-like cells. Appearance of beating colonies from human iPSCs was increased approximately 4.3 times by addition of CSA at mesoderm stage. CSA-expanded human iPSC-derived cardiomyocytes showed various cardiac marker expressions, synchronized calcium transients, cardiomyocyte-like action potentials, pharmacological reactions, and ultra-structural features as cardiomyocytes. These results provide a technological basis to obtain functional cardiomyocytes from iPSCs.
authors.affiliation: Laboratory of Stem Cell Differentiation, Stem Cell Research Center, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan.
authors.forename: Masataka
authors.initials: M
authors.lastname: Fujiwara
authors.forename: Peishi
authors.initials: P
authors.lastname: Yan
authors.forename: Tomomi G
authors.initials: TG
authors.lastname: Otsuji
authors.forename: Genta
authors.initials: G
authors.lastname: Narazaki
authors.forename: Hideki
authors.initials: H
authors.lastname: Uosaki
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Fukushima
authors.forename: Koichiro
authors.initials: K
authors.lastname: Kuwahara
authors.forename: Masaki
authors.initials: M
authors.lastname: Harada
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Matsuda
authors.forename: Satoshi
authors.initials: S
authors.lastname: Matsuoka
authors.forename: Keisuke
authors.initials: K
authors.lastname: Okita
authors.forename: Kazutoshi
authors.initials: K
authors.lastname: Takahashi
authors.forename: Masato
authors.initials: M
authors.lastname: Nakagawa
authors.forename: Tadashi
authors.initials: T
authors.lastname: Ikeda
authors.forename: Ryuzo
authors.initials: R
authors.lastname: Sakata
authors.forename: Christine L
authors.initials: CL
authors.lastname: Mummery
authors.forename: Norio
authors.initials: N
authors.lastname: Nakatsuji
authors.forename: Shinya
authors.initials: S
authors.lastname: Yamanaka
authors.forename: Kazuwa
authors.initials: K
authors.lastname: Nakao
authors.forename: Jun K
authors.initials: JK
authors.lastname: Yamashita
mesh_terms: D000328:Adult; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects* / Q000502:physiology; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D018920:Coculture Techniques; D016572:Cyclosporine / Q000494:pharmacology*; D055724:Electrophysiological Phenomena / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects* / Q000502:physiology; D051379:Mice; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000502:physiology; D013045:Species Specificity; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D004791:Enzyme Inhibitors; D016572:Cyclosporine