Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (10): 1786-1790.doi: 10.3969/j.issn.1673-8225.2012.10.017

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Preparation of CF-1 mouse embryonic fibroblast feeder layers

Lu Hai1, Zhang Zhi1, Zhao Yi-chao1, Rao Xiao-hui1,Jiang Jin-qun2, Meng Bin3, Ai Min4, Pan Ming-xin1, Gao Yi1   

  1. 1Second Department of Hepatobiliary Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou  510280, Guangdong Province, China; 2Anhui Bangbu Medical College, Bangbu  233030, Anhui Province, China; 3Medical College of Anhui University of Science and Technology, Huainan  232001, Anhui Province, China; 4the 305 Hospital of Chinese PLA, Beijing  100017, China
  • Received:2011-12-16 Revised:2012-01-22 Online:2012-03-04 Published:2012-03-04
  • Contact: author: Gao Yi, Chief physician, Professor, Doctoral supervisor, Second Department of Hepatobiliary Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510280, Guangdong Province, China gaoyi6146@163.com
  • About author:Lu Hai, Studying for master’s degree, Physician, Second Department of Hepatobiliary Surgery, Guangzhou 510280, Guangdong Province, China hysa1985@yahoo.com.cn
  • Supported by:

    the Science Research Program of Military Medicine and Hygiene, No.08Z017*; National High-Tech Research and Development Program of China (863 Program), No.2006AA02A141*

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Abstract:

BACKGROUND: Mouse embryo fibroblasts as the feeder layer for growth of stem cells exhibit better effects than the circumstance without feeder layer because they can secrete some factors which can promote the growth of stem cells and inhibit the differentiation of stem cells.
OBJECTIVE: To establish the optimal method for isolation and culture of CF-1 mouse embryo fibroblast feeder layer and investigate the optimal concentration of mitomycin C in inhibiting the proliferation of fibroblasts and the processing time.
METHODS: Fetal mice were taken from CF-1 mice at pregnancy days 10-15. Primary fibroblasts were isolated and treated with different concentrations (5, 10, 15, 20 mg/L) of mitomycin C for different time periods (1, 1.5, 2, 2.5, 3 hours) to prepare feeder layers. Cell proliferation was observed. Human induced pluripotent stem cells or embryonic stem cells were cultured on mitomycin C-treated feeder layer and the growth of cell colonies was observed.
RESULTS AND CONCLUSION: The optimal fetal age for preparing CF-1 mouse embryo fibroblast feeder layer was 13.0-14.0 days. Treatment with 10 mg mitomycin C for 2.5 hours showed optimal effects on inhibiting the proliferation of CF-1 mouse embryo fibroblasts and fibroblast feeder layer could maintain 5-7 days. Induced pluripotent stem cells and embryonic stem cells can develop into typical “bird nest”-shaped colony of stem cells after treatment with 10 mg/L mitomycin C for 2.5 hours.

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