Abstract: In order to study the interaction mechanism between maternal and conceptus of sheep,an in vitro implantation model was established by establishing and improving the in vitro culture technology of primary sheep endometrial epithelial cells.Endometrial epithelial cells were isolated and cultured by tissue block method.The growth of endometrial epithelial cells was observed,and the effects of different EGF concentrations on the proliferation of endometrial epithelial cells were compared.The results showed that after 1-2 days of tissue culture,the endometrial epithelial cells migrated from the peri-tissue,and it took about 9-12 days to grow the 60 mm culture dish.After purification by differential digestion,the purity of the endometrial epithelial cells of F1 generation sheep could reach more than 90%,which indicated that the obtained cells could be used in subsequent experiments.F2 cells were cultured at different EGF concentrations (0,12.5,25,50,75 and 100 ng/mL) for 144 and 168 hours.The D_{450 nm} values were extremely significant higher in 12.5,25 and 100 ng/mL EGF treatment group (P<0.01),and significantly higher in 50 ng/mL EGF treatment group (P<0.05) compared with control group at 144 h.At 168 h,the D_{450 nm} values were significantly higher in 12.5 ng/mL EGF treatment group than those in control group (P<0.05).Therefore,at 12.5 ng/mL EGF concentration,the effect was the best.At 0 and 12.5 ng/mL EGF concentrations,F3 cells were cultured for different time (24,48,72,96,120,144,168,192 and 216 h).The D_{450 nm} values were significantly different at 144 h (P<0.05),and extremely significant at 168 h (P<0.01).Consequently,the culture medium with 12.5 ng/mL EGF would obtain the endometrial epithelial cells more efficiently and rapidly.

Key words: sheep; endometrial epithelial cells; EGF; immunofluorescence staining