A novel efficient feeder-free culture system for the derivation of human induced pluripotent stem cells

被引:485
作者
Nakagawa, Masato [1 ]
Taniguchi, Yukimasa [2 ]
Senda, Sho [3 ]
Takizawa, Nanako [1 ]
Ichisaka, Tomoko [1 ]
Asano, Kanako [1 ]
Morizane, Asuka [1 ]
Doi, Daisuke [1 ]
Takahashi, Jun [1 ]
Nishizawa, Masatoshi [1 ]
Yoshida, Yoshinori [1 ]
Toyoda, Taro [1 ]
Osafune, Kenji [1 ]
Sekiguchi, Kiyotoshi [2 ]
Yamanaka, Shinya [1 ,4 ]
机构
[1] Kyoto Univ, Ctr iPS Cell Res & Applicat CiRA, Kyoto 6068507, Japan
[2] Osaka Univ, Inst Prot Res, Osaka 5650871, Japan
[3] Ajinomoto CO Inc, Inst Innovat, Kawasaki, Kanagawa 2108681, Japan
[4] Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA
来源
SCIENTIFIC REPORTS | 2014年 / 4卷
基金
日本学术振兴会;
关键词
HUMAN FIBROBLASTS; SELF-RENEWAL; DIFFERENTIATION; GENERATION; BANKING; GROWTH; LINES;
D O I
10.1038/srep03594
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
In order to apply human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) to regenerative medicine, the cells should be produced under restricted conditions conforming to GMP guidelines. Since the conventional culture system has some issues that need to be addressed to achieve this goal, we developed a novel culture system. We found that recombinant laminin-511 E8 fragments are useful matrices for maintaining hESCs and hiPSCs when used in combination with a completely xeno-free (Xf) medium, StemFit (TM). Using this system, hESCs and hiPSCs can be easily and stably passaged by dissociating the cells into single cells for long periods, without any karyotype abnormalities. Human iPSCs could be generated under feeder-free (Ff) and Xf culture systems from human primary fibroblasts and blood cells, and they possessed differentiation abilities. These results indicate that hiPSCs can be generated and maintained under this novel Ff and Xf culture system.
引用
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页数:7
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