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Open Access Highly Accessed Methodology article

Establishment of automated culture system for murine induced pluripotent stem cells

Hiroyuki Koike1, Koji Kubota2, Keisuke Sekine1, Takanori Takebe1, Rie Ouchi1, Yun-Wen Zheng1, Yasuharu Ueno1, Naoki Tanigawa2 and Hideki Taniguchi1*

Author affiliations

1 Department of Regenerative Medicine, Graduate School of Medicine, Yokohama City University, 3-9 Fukuura, Kanazawa-ku, Yokohama, Kanagawa 236-0004, Japan

2 Chiyoda Corporation, Minatomirai Grand Central Tower, 4-6-2 Minatomirai, Nishi-ku, Yokohama, Kanagawa 220-8765, Japan

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Citation and License

BMC Biotechnology 2012, 12:81  doi:10.1186/1472-6750-12-81

Published: 5 November 2012

Abstract

Background

Induced pluripotent stem (iPS) cells can differentiate into any cell type, which makes them an attractive resource in fields such as regenerative medicine, drug screening, or in vitro toxicology. The most important prerequisite for these industrial applications is stable supply and uniform quality of iPS cells. Variation in quality largely results from differences in handling skills between operators in laboratories. To minimize these differences, establishment of an automated iPS cell culture system is necessary.

Results

We developed a standardized mouse iPS cell maintenance culture, using an automated cell culture system housed in a CO2 incubator commonly used in many laboratories. The iPS cells propagated in a chamber uniquely designed for automated culture and showed specific colony morphology, as for manual culture. A cell detachment device in the system passaged iPS cells automatically by dispersing colonies to single cells. In addition, iPS cells were passaged without any change in colony morphology or expression of undifferentiated stem cell markers during the 4 weeks of automated culture.

Conclusions

Our results show that use of this compact, automated cell culture system facilitates stable iPS cell culture without obvious effects on iPS cell pluripotency or colony-forming ability. The feasibility of iPS cell culture automation may greatly facilitate the use of this versatile cell source for a variety of biomedical applications.

Keywords:
Induced pluripotent stem (iPS) cell; Automated cell culture system (ACCS); CO2 incubator-scale; Pluripotency