Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (45): 8429-8433.doi: 10.3969/j.issn.1673-8225.2011.45.015

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Optimization of separation methods and culture system of chicken embryonic stem cells in vitro

Zhang Man-yu1, Chen Zhi-cheng2, Ji Hui-qin2, Chen Sheng-feng2, Liu Ben-jie1, Deng Xian-bo1, Wang Bing-yun2, Yuan Li3, Jiang Qing-yan4   

  1. 1Veterinary College, South China Agricultural University, Guangzhou  510642, Guangdong Province, China
    2Veterinary Department, College of Life Science, Foshan University, Foshan  528231, Guangdong Province, China
    3Department of Biomedicine, School of Life Science, Xiamen University, Xiamen  361005, Fujian Province, China
    4College of Animal Science, South China Agricultural University, Guangzhou  510642, Guangdong Province, China
  • Received:2011-05-06 Revised:2011-06-18 Online:2011-11-05 Published:2011-11-05
  • Contact: Deng Xian-bo, Master, Associate professor, Master’s supervisor, Veterinary College, South China Agricultural University, Guangzhou 510642, Guangdong Province, China xbdeng@scau.edu.cn Correspondence to: Wang Bing-yun, Doctor, Professor, Master’s supervisor, Veterinary Department, College of Life Science, Foshan University, Foshan 528231, Guangdong Province, China bywang63@163.com
  • About author:Zhang Man-yu★, Studying for master’s degree, Veterinary College, South China Agricultural University, Guangzhou 510642, Guangdong Province, China Linyizhangmanyu @163.com
  • Supported by:

    the National 973 Program of China, No. 2009CB941600*; the National Natural Science Foundation of China, No. 31072101*

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Abstract:

BACKGROUND: Embryonic stem cells are undifferentiated permanent cell line derived from inner cell mass cells and primordial germ cells of animal’s early embryos. Chicken embryonic stem cells are derived from the blastodermal of a X-stage embryo.
OBJECTIVE: To optim the separation method and in vitro cultural system of chicken embryonic stem cells.
METHODS: The X-stage chicken embryos were isolated by using a small square of ?lter paper with a hole punched in the center, and the blastodermal cells were isolated by using the hair loop. STO cells were used to make feeder layer; at the same time, BRL-CM and cytokine were also used for chicken embryonic stem cells in vitro cultural system.
RESULTS AND CONCLUSION: The filter paper loop and the hair loop could obtain complete the blastoderm, and the successful percentage was 75%-85%. The colony formation rate was about 50%. After culture in the BRL-CM + feeder layer + cytokine culture system, the passage of CES cells is the seventh generation; BRL-CM + feeder layer + cytokines, cultured chicken embryonic stem cells could passage to the 25th generation. Isolated chicken embryonic stem cells were in an undifferentiated state detected by alkaline phosphatase staining and SSEA-1 staining. The findings indicate that this experiment not only optimized the isolation method of chicken embryonic stem cells to obtain complete and pure embryos, but also further improved the in vitro culture system of chicken embryonic stem cells.

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