|Cancer||Cell death||Cell cycle||Cytoskeleton||Exo/endocytosis||Differentiation||Division||Organelles||Signalling||Stem cells||Trafficking|
Cell Biology International (2009) 33, 12681273 (Printed in Great Britain)
Role of MEF feeder cells in direct reprogramming of mousetail-tip fibroblasts
Mengfei Chenab, Xuerong Suna, Ruzhang Jianga, Wenjuan Shenc, Xiufeng Zhonga, Bingqian Liua, Ying Qia, Bing Huanga, Andy Peng Xiangb and Jian Gea*
aState Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, 54 Xian Lie Nan Road, Guangzhou 510060, China
bCenter for Stem Cell Biology and Tissue Engineering, Sun Yat-sen University, Guangzhou 510080, China
cDepartment of Pathophysiology, Medical College of Jinan University, Guangzhou 510632, China
Pluripotent stem cells can be induced from somatic cells by the transcription factors Oct3/4, Sox2, c-Myc and Klf4 when co-cultured with mouse embryonic fibroblast (MEF) feeder cells. To date, the role of the feeder cells in the reprogramming process remains unclear. In this study, using a comparative analysis, we demonstrated that MEF feeder cells did not accelerate reprogramming or increase the frequency of induced pluripotent stem (iPS) cell colonies. However, feeder conditions did improve the growth of primary iPS colonies and were necessary for passaging the primary colonies after reprogramming was achieved. We further developed a feeder-free culture system for supporting iPS growth and sustaining pluripotency by adding bFGF and activin A (bFA) to the medium. These data will facilitate the generation of human iPS cells without animal feeders for regenerative medicine.
Key words: Induced pluripotent stem cells, Embryonic stem cells, Reprogramming, fibroblasts, MEF feeder cells, Feeder-free.
*Corresponding author. Tel.: +86 20 87331374; fax: +86 20 87333271.
Received 10 January 2009/18 May 2009; accepted 3 June 2009doi:10.1016/j.cellbi.2009.06.004