Induction and Enhancement of Cardiac Cell Differentiation from Mouse and Human Induced Pluripotent Stem Cells with Cyclosporin-A

被引：93

作者：

Fujiwara, Masataka ^{[1
,2
]}

Yan, Peishi ^{[1
,3
]}

Otsuji, Tomomi G. ^{[4
,5
]}

Narazaki, Genta ^{[1
,6
]}

Uosaki, Hideki ^{[1
,6
]}

Fukushima, Hiroyuki ^{[1
,6
]}

Kuwahara, Koichiro ^{[2
]}

Harada, Masaki ^{[2
]}

Matsuda, Hiroyuki ^{[7
]}

Matsuoka, Satoshi ^{[7
]}

Okita, Keisuke ^{[8
]}

Takahashi, Kazutoshi ^{[8
]}

Nakagawa, Masato ^{[8
]}

Ikeda, Tadashi ^{[3
]}

Sakata, Ryuzo ^{[3
]}

Mummery, Christine L. ^{[9
]}

Nakatsuji, Norio ^{[10
,11
]}

Yamanaka, Shinya ^{[8
,12
]}

Nakao, Kazuwa ^{[2
]}

Yamashita, Jun K. ^{[1
,6
]}

机构：

[1] Kyoto Univ, Inst Frontier Med Sci, Stem Cell Res Ctr, Lab Stem Cell Differentiat, Kyoto, Japan

[2] Kyoto Univ, Dept Med & Clin Sci, Grad Sch Med, Kyoto, Japan

[3] Kyoto Univ, Dept Cardiovasc Surg, Grad Sch Med, Kyoto, Japan

[4] Stem Cell & Drug Discovery Inst, Kyoto, Japan

[5] Kyoto Univ, Inst Frontier Med Sci, Stem Cell Res Ctr, Lab Embryon Stem Cell Res, Kyoto, Japan

[6] Kyoto Univ, Ctr iPS Cell Res & Applicat CiRA, Dept Cell Growth & Differentiat, Kyoto, Japan

[7] Kyoto Univ, Dept Physiol & Biophys, Grad Sch Med, Kyoto, Japan

[8] Kyoto Univ, Ctr iPS Cell Res & Applicat CiRA, Dept Reprogramming Sci, Kyoto, Japan

[9] Leiden Univ, Med Ctr, Dept Anat & Embryol, Leiden, Netherlands

[10] Kyoto Univ, Inst Frontier Med Sci, Dept Dev & Differentiat, Kyoto, Japan

[11] Kyoto Univ, Inst Integrated Cell Mat Sci iCeMS, Kyoto, Japan

[12] Gladstone Inst Cardiovasc Dis, San Francisco, CA USA

来源：

PLOS ONE | 2011年 / 6卷 / 02期

关键词：

CARDIOVASCULAR PROGENITOR CELLS; SMOOTH-MUSCLE; GENERATION; CARDIOMYOCYTES; FIBROBLASTS; MYOCYTES;

D O I：

10.1371/journal.pone.0016734

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页数：10

共 38 条

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