Ascorbic acid enhances the cardiac differentiation of induced pluripotent stem cells through promoting the proliferation of cardiac progenitor cells  

作　　者：Nan Cao Zumei Liu Zhongyan Chen Jia Wang Taotao Chen Xiaoyang Zhao Yu Ma Lianju Qin Jiuhong Kang Bin Wei Liu Wang Ying Jin Huang-Tian Yang 

机构地区：[1]Key Laboratory of Stem Cell Biology, Institute of Health Sciences, Shanghai Institutes for Biological Sciences （SIBS）, Chinese Academy of Sciences （CAS） ＆ Shanghai Jiao Tong University School of Medicine （SJTUSM）, Shanghai 200025, China [2]Laborato- ry of Molecular Cell Biology, Institute of Biochemistry and Cell Biology, SIBS, CAS, Shanghai 200031, China [3]State Key Labora- tory of Reproductive Biology, Institute of Zoology, CAS, Beijing I00101, China [4]Shanghai Stem Cell Institute, SJTUSM, Shanghai 200025, China

出　　处：《Cell Research》2012年第1期219-236,共18页细胞研究（英文版）

基　　金：This study was supported by grants from the National Basic Research Program of China （2011 CB965300, 2009CB941100, 2010CB945600）, the National Natural Science Foundation of China （31030050）, Strategic Priority Research Program of CAS （XDA01000000）, Science and Technology Committee of Shanghai Municipality （08DJ1400501）, National Science and Technology Project of China （2012ZX09501-001-001）, and Sanofi-Aventis Re- cherche ＆ D6veloppement-SIBS funding. We thank Dr Duanqing Pei （Guangzhou Institutes of Biomedicine and Health, China） for kindly providing the miPSC lines iPS-C5 and iPS-C12.

摘　　要：Generation of induced pluripotent stem cells （iPSCs） has opened new avenues for the investigation of heart diseases, drug screening and potential autologous cardiac regeneration. However, their application is hampered by inefficient cardiac differentiation, high interline variability, and poor maturation of iPSC-derived cardiomyocytes （iPS-CMs）. To identify efficient inducers for cardiac differentiation and maturation of iPSCs and elucidate the mechanisms, we systematically screened sixteen cardiomyocyte inducers on various mnrine （m） iPSCs and found that only ascorbic acid （AA） consistently and robustly enhanced the cardiac differentiation of eleven lines including eight without spontaneous cardiogenic potential. We then optimized the treatment conditions and demonstrated that differentiation day 2-6, a period for the specification of cardiac progenitor cells （CPCs）, was a critical time for AA to take effect. This was further confirmed by the fact that AA increased the expression of cardiovascular but not mesodermal markers. Noteworthily, AA treatment led to approximately 7.3-fold （miPSCs） and 30.2-fold （human iPSCs） augment in the yield of iPS-CMs. Such effect was attributed to a specific increase in the proliferation of CPCs via the MEK-ERK1/2 pathway by promoting collagen synthesis. In addition, AA-induced cardiomyocytes showed better sarcomeric organization and enhanced responses of action potentials and calcium transients to p-adrenergic and muscarinic stimulations. These findings demonstrate that AA is a suitable cardiomyocyte inducer for iPSCs to improve cardiac differentiation and maturation simply, universally, and efficiently. These findings also highlight the importance of stimulating CPC proliferation by manipulating extracellular microenvironment in guiding cardiac differentiation of the pluripotent stem cells.

关 键 词：induced pluripotent stem cells ascorbic acid CARDIOMYOCYTES cardiac progenitor cells 

分 类 号：Q813[生物学—生物工程] Q813.11