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

In a joint research project with Osaka University and Ajinomoto Co., Inc., a research team led by Masato Nakagawa (lecturer at Kyoto University, CiRA) and Professor Shinya Yamanaka (Kyoto University, CiRA) has developed a new method for generation and maintenance culture of induced pluripotent stem cells (iPS cells) that are suitable for use in cell transplantation therapy.

In order to use human iPS/ES cells in regenerative medicine, the cells must be prepared using methods compliant with GMP (Good Manufacturing Practice). However, the methods in use up till now have used feeder cells to culture iPS/ES cells, which involves complicated procedures, and have employed culture medium containing serum and numerous other animal-derived constituents, which made it difficult for them to comply with GMP.

In the method discovered by Dr. Nakagawa and his research team, feeder cells were replaced with recombinant laminin 511-E8 fragments (iMatrix-511, Nippi), which were developed by the research team of Professor Kiyotoshi Sekiguchi of Osaka University Institute for Protein Research and manufactured to GMP standard by Nippi, Incorporated. The cells were maintained in a culture medium free of animal-derived constituents (xeno-free) which was developed jointly with Ajinomoto. Using this method facilitates the handling of human iPS/ES cells and makes it possible to perform stable and long-term serial passage culture without generating chromosomal abnormalities.

These iPS cells generated from human skin or blood cells with no use of feeder cells (feeder-free) and cultured in xeno-free medium (StemFit, AJINOMOTO) were transplanted into immunodeficient mice. Teratoma formation was observed and the cells were confirmed to be capable of differentiating into the cells of all three germ layers. The researchers also succeeded in inducing the iPS cells thus produced to differentiate into dopamine-producing cells, insulin-producing cells, and blood cells. 

These findings demonstrate that generation and maintenance culture of human iPS cells is possible using this newly developed feeder-free and xeno-free culture system. This method will not only make it possible to generate iPS cells of a quality ideally suited to cell transplantation into humans, but should also prove useful in areas such as drug discovery, toxicity testing, and disease modelling. The findings of the research will be published in the British scientific journal Scientific Reports on January 8.

The protocol for our iPSC-culture system is available from CiRA web page.

Now you can try StemFit medium as a free sample. Please contact us (nakagawa-g@cira.kyoto-u.ac.jp)

Masato Nakagawa , Yukimasa Taniguchi , Sho Senda , Nanako Takizawa , Tomoko Ichisaka , Kanako Asano, Asuka Morizane, Daisuke Doi, Jun Takahashi, Masatoshi Nishizawa, Yoshinori Yoshida, Taro Toyoda, Kenji Osafune, Kiyotoshi Sekiguchi, and Shinya Yamanaka

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

SCIENTIFIC REPORTS | 4 : 3594 | DOI: 10.1038/ srep03594 | 2014 |

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