Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (10): 1780-1784.doi: 10.3969/j.issn.1673-8225.2010.10.014

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Screening of optimal embryonic time for in vitro separation and culture of Kunming mouse embryonic stem cells: Comparison among 2.5, 3.5, and 4.5 pregnant days

Zhang Hai-feng1, Shan Wei2, Qin Shu-jian2   

  1. 1Department of Anatomy, Xuzhou Medical College, Xuzhou   221004, Jiangsu Province, China;
    2Department of Anatomy, Liaoning Medical University, Jinzhou   121001, Liaoning Province, China
  • Online:2010-03-05 Published:2010-03-05
  • About author:Zhang Hai-feng, Master, Department of Anatomy, Xuzhou Medical College, Xuzhou 221004, Jiangsu Province, China pearlbead2004@ sina.com
  • Supported by:

    the Natural Science Foundation of Liaoning Province, No. 20032142*;
    the Project Foundation of Xuzhou Medical College, No. 08KJ19*

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Abstract:

BACKGROUND: Genotype of Kunming mice was similar to human population, thus an establishment of embryonic stem cell line is beneficial for research of transgenic animal. However, the best time to collect embryo has been less reported yet.

OBJECTIVE: To find the best time to collect embryos from Kunming mice.

METHODS: The embryos were collected from mother mice of 2.5, 3.5, and 4.5 pregnant days. Microscope was used to evaluate the growth condition of embryos, embryo attaching rate (A/C), inner cell mass (ICM) growing rate (I/C), embryonic stem cells (ESCs) clone growing rate (P1/C) and ESCs subclone growing rate (P2/C). The cells were then stained with alkaline phosphatase.

RESULTS AND CONCLUSION: Most of the 2.5-pregnant-day embryos were 16-cell-phase embryos. The 3.5-pregnant-day embryos were morulas while the 4.5-pregnant-day embryos were blastulas. There were no significant differences in A/C, I/C, P1/C, P2/C between 2.5 and 3.5 pregnant days (P > 0.05). The 4.5-pregnant-day indicators mentioned above were significantly greater than those two groups; therefore, 4.5-pregnant-day embryos were the best source to culture, clone, isolate and passage ESCs.

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