Study on Mouse Oviductal Epithelial Primary Cell Culture and Its Purification

To obtain a good method of primary culture of oviductal epithelial cells(OECs) and find a way of purification based on the model of mouse.Two methods of primary culture of OECs,tissue explants and enzymic digestion were used.The latter one was divided into 4 treatments that all were disposaled under 37 ℃.The diced tissue blocks were digested for 5 min,10 min and 20 min with 2.5 g/L trypsin+0.4 g/L EDTA in treatment 1,for 35 min,50 min and 75 min with 0.5 g/L trypsin+0.08 g/L EDTA in treatment 2,for 60 min,90 min and 240 min with 0.3 g/L collagenaseⅠ in treatment 3,for 60 min and 150 min with 0.3 g/L collagenaseⅠ and 2.5 g/L trypsin+0.4 g/L EDTA(2∶1) in treatment,4 respectively.As for purification of OECs,the ways of different speed of cell attachment(DSCA) and re-DSCA were adopted in primary culture.On the other hand,two-step digestion was adopted to purify OECs in tissue explant subculture.Fibroblasts were mixed with OECs in primary cell culture.Furthermore,fibroblasts were superior to growth,which greatly affected subculture.As to enzymic digestion,treatment 1 had a result with a low attachment efficiency;Treatment 2 gained relatively good results.Treatment 3 only in digestion for 240 min obtained ideal result.It was better in digestion for 150 min in treatment 4.In primary culture,more OECs with less fibroblasts were gained by adopting re-DSCA.The effect was good by adopting two-step digestion in subculture.It is advised that enzyme digestion as well as re-DSCA during process of primary cell culture and two-step digestion during process of passage could be adopted to purify OECs successfully.