In vitro culture of mouse endometrium

    TAN Dongª²Mei1, TAN Yi1, ZHAO Jie2, AI Ling2, WANG Zhiª²Biao2

    1Laboratory Animal Center, 2Ultrasound Engineering Institute of Medical Science, Chongqing University of Medical Science, Chongqing 400016, China

    ¡¾Abstract¡¿ AIM: To develop an in vitro culture model of mouse endometrium. METHODS: Mouse endometrium squeezed from the uterus horns were cultured at the gasª²liquid interface between F12/DMEM (1¡Ã1) including 200 mL/L FBS, 17.5 nmol/L insulin, 63.5 nmol/L progesterone, 7.14 nmol/L estradiol, and 50 mL/L CO2 plus 950 mL/L air. The effects of 20 ¦Ìg/L epidermal growth factor (EGF) and those of mixed gas containing 50 mL/L CO2 plus 950 mL/L O2 on the maintenance of the cultured endometrium were compared. The necrosis rate of the samples was measured after hematoxylinª²eosin staining by using the image analysis system. The expressions of leukemia inhibitory factor (LIF) and heparinª²binding epidermal growth factorª²like growth factor (HBª²EGF) in the mouse endometrium were detected by immunohistochemistry to determine whether the endometrium cultured for 24 h was still receptive to the blastocysts. RESULTS: The necrosis rate (%) of the endometrium in the three groups was 19.40¡À3.38, 16.41¡À3.40 and 13.71¡À2.89 respectively, showing a significant difference (P=0.000). The positively rates of LIF and HBª²EGF proteins in both cultured and controlled mouse endometrium were 100% unexceptionally and there was no significant difference in the staining intensity for each marker before and after the culture (P=0.237, 0.224). CONCLUSION: The culture condition: F12/DMEM (1¡Ã1), 200 mL/L FBS, 17.5 nmol/L insulin, 63.5 nmol/L progesterone, 7.14 nmol/L estradiol, 20 ¦Ìg/L EGF and 50 mL/L CO2 plus 950 mL/L O2, proves to be most optimal for the growth of mouse endometrium in vitro. The endometrium cultured for 24 hours is still receptive to the blastocysts. ......