Abstract:

BACKGROUND: A lot of methods about the isolation, culture and induced decidualization in vitro of endometrial stromal cells have been established in China, but there is a problem about drawn difficulties of uterine specimens.
OBJECTIVE: To establish the method of isolation, culture and induced decidualization in vitro of mouse endometrial stromal cells, and to provide a simple and effective model for further study about the mechanism of endometrial decidualization.
METHODS: The trypsin and collagenase Ⅱ digestion and pasted wall were used to isolate and purify endometrial stromal cells of the mouse that pregnancy for 4 days; stromal cells were identified by immunocytochemistry; endometrial stromal cells treated by estradiol and progesterone were induced the decidualization, and the decidual markers were checked by hematoxylin nuclear staining, real-time quantitative PCR and western blot.
RESULTS AND CONCLUSION: The survival rate of endometrial stromal cells was (90.1± 2.3)%. Immunocytochemistry staining results showed that separated and purified uterine stromal cells were stained positive with Vimentin, and negative with Cytokeratin, the purities of cells were (92.3±2.4)%. Hematoxylin staining showed that endometrial stromal cells were multinucleated after treated with estradiol and progesterone for 48 and 72 hours. The decidualization prolactin-related protein, cyclin D3, progesterone receptor mRNA of decidual markers were significantly increased with the incubation time prolonged (P < 0.05). Highly purified stromal cells of endometrium can be obtained by enzymolysis and pasted wall purification. Endometrial stromal cells treated by estradiol and progesterone can be induced to the decidualization in vitro.