The isolation and characterization of a telomerase immortalized goat trophoblast cell line
Trophoblast cells play vital roles in the processes of embryonic implantation and placentation.Many toxicological compounds can induce the malfunction of trophoblast cells, resulting in implantation failure or early embryonic loss.The finite lifespan of primary trophoblast cells limits investigation of the long-term effects of some toxicological compounds on trophoblast cells in vitro.In this study, primary goat trophoblast cells were purified by density gradient centrifugation and specific immuno-affinity purification.Then, the purified cells were immortalized through transfection of a plasmid containing the human telomerase reverse transcriptase (hTERT) gene.hTERT-transfected goat trophoblast cells (hTERT-GTCs) could steadily express hTERT gene and exhibit higher telomerase activity, and persistently proliferate without any signs of senescence up to 50 passages.The immortalized goat trophoblast cells still possessed the basic and key properties of normal primary goat trophoblast cells to express the specific intracellular marker cytokeratin 7 (CK-7) and secrete chorionic gonadotrophin β-subunit (CG-β) and placental lactogen (PL).Further studies showed that the immortalized goat trophoblast cells expressed vimentin and non-classical MHC class I antigen and exhibited invasive phenotype, suggesting that the immortalized goat trophoblasts resembled human extravillous trophoblasts.In addition, this cell line did not show neoplastic transformation either in vivo or in vitro.We concluded the immortalized goat trophoblast cells by hTERT transfeetion retained the basic and key characteristics of primary trophoblast cells and may provide a useful model to study the effects of some toxicological compounds on trophoblast cells.
Goat trophoblast cells hTERT Telomerase Immortalization
Feng Dong Yong Huang Wei Li Xiaomin Zhao Wenlong Zhang Qian Du Hongling Zhang Xiangjun Song Dewen Tong
College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100,P.R.China
国内会议
南京
英文
31-42
2013-10-01（万方平台首次上网日期，不代表论文的发表时间）