Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (45): 8496-8500.doi: 10.3969/j.issn.2095-4344.2012.45.025

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Cloning efficiency of human endometrial cells in vitro

Ma Ying, He Yuan-li   

  1. Department of Gynecology and Obstetrics, Zhujiang Hospital, Southern Medical University,Guangzhou 510280, Guangdong Province, China
  • Received:2012-01-08 Revised:2012-03-28 Online:2012-11-04 Published:2012-11-04
  • About author:Ma Ying☆, Doctor, Attending physician, Department of Gynecology and Obstetrics, Zhujiang Hospital, Southern Medical University, Guangzhou 510280, Guangdong Province, China xxbcxh@sohu.com

Abstract:

BACKGROUND: Colony forming ability is an important characteristic of stem cells. Many studies have shown that the endometrium has monoclonal characteristics.
OBJECTIVE: To detect the cloning efficiency of human endometrial cells in vitro.
METHODS: Human endometrial cells were isolated and dissociated mechanically and enzymatically from human endometrium. Stromal and epithelial cells were separated by two series of filters. The cell morphology and growth characteristics were observed and the expressions of vimentin and keratin were detected. Flow cytometry was used to identify the expression of CD133, CD34, CD45, CD90, CD73 and CD29 markers in endometrial cells. The clone growth characteristics were observed, the size of clones was measured and the rate of cloning efficiency was calculated. Meanwhile, human uterine muscle cell was used as a control.
RESULTS AND CONCLUSION: After primary cultured for 12 days, glandular epithelial cells and stromal cells were divided into large and small clones by size and growth characteristics. The gland clone was positive for keratin and the stromal clone was positive for vimentin by immunohistochemistry. Results from flow cytometry showed that endometrial cells were positive for CD29, CD90 and CD73 but negative for CD34, CD45 and CD133. There was significant difference of the sizes and cloning effiency of the large and little gland clones between glandular epithelial cells and stromal cells (P < 0.05). Human uterine smooth muscle cells were primary cultured by limiting dilution method. The endometrial cells can form landular epithelial cells and stromal cells in vitro which included large and small clones. The proliferation efficiency of the large gland clone was higher than that of the small gland clone. These cells may be the endometrial stem cells.

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