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In Vitro Culture of Trophoblast Giant Cells from Bovine Placentomes
Yan Ren, Chuang-Fu Chen*, Jun Qiao, Yong Wang1, Qian Guo, Hui Zhang, Peng-Yan Wang
Laboratory of Xinjiang Endemic and Ethnic Disease, College of Animal Science & Technology, Shihezi University, Shihezi 832003, China; 1College of Life Science, Shihezi University, Shihezi 832003, China
Abstract: To establish a convenient method of purification and culture bovine trophoblast cells. Healthy placental tissues between 45 and 60 days of gestation were obtained and digested by collagen I， and then the cells were randomly divided into two groups. The cells from one group were purified by different speeding adherence. Finally， the cells were cultured in DMEM medium including 15% FCS and 5% trophoblast grouth supplement (TGS). The cellular morphology and vitality were observed by inverted phase contrast microscope and trypan blue staining. The nucleus were observed by Hoechst 33342. Immunocytochemistry was used to examine the expression of cytokeratin and vimentin. The results showed that the method of purification had significant effect. The livability was above 90% and typical two nucleus were stained blue. The isolated binucleate trophoblast giant cell (TGC) were positive for cytokeratin and negative for vimentin throughout the duration of the experiment. The purified cells contained 95% TGC and unpurified cells contained only 45%-50% TGC. Cells could passage four generations and survived for sixty days. We had developed an efficient culture system for differentiated TGC enabling future research in this field.
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