| To estalish a stable,convenient and high purified method used for obtaining placental trophoblast cells in yak, the present experiment was carried out by use of co-digestive solution to digest the fetal placental tissues in the age of 8 to 10 weeks, and single trophoblast cell suspension was made. The trophoblast cells was then isolated and purified.The optimal concentration of FBS and optimal PH were determined for culturing the trophoblast cells; meanwhile, the general morphology and nuclear characteristics of the trophoblast cells, and the expression of cytokeratin7 (CK 7) and vimentin (Vim) were also examined. The results obtained were as the follow.(1)The collected placenta tissue samplesshould betreated within 24h when it was keep in Hank's solution with a temperature of4℃, A good way to separate trophoblast cells was the conbined digestion method used the trypsin and collagenase in a proportion of 1:1 separated, basically trophderm cell and other impurity cells would be roughly separated though a 400 item of strainer in the percoll separating medium.(2)Thecultivating condition of 20%FBSDMEM/F12 medium with a pH6.8~7.0 was most suitable for primary culture of the placenta trophoblast cell, while the condition of 10%FBSDMEM/F12 medium with a pH6.8~7.0 was suitable for subculture of the placenta trophoblast cell.(3) It was determined by perspective microscope that after 5 times subculturing the trophoblast cells shows epithelial-like shape and grew in a schistic spreading way.The immunohistochemistry staining showed that the cytokeratin chromosome was positive, while the vimentin chromosomenegative.The positive cells rate amounted to 90% and the cell viability evaluated was more than 95% by trpan blue.The researchresults indicated that a stable,efficient and high purified method used for subcultivating the trophoblast cells in yak could be established by used of the digestion and discharge purification in a condition of 10%FBSDMEM/F12 medium with a pH6.8~7.0.
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