The isolation and characterization of a telomerase immortalized goat trophoblast cell line
The isolation and characterization of a telomerase immortalized goat trophoblast cell line
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端粒酶永生化山羊滋养层细胞系的分离和鉴定
DOI：
10.1016/j.placenta.2013.09.009
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发表时间：
2013-12-01
期刊：
影响因子：
3.8
通讯作者：
Tong, D.
中科院分区：
文献类型：
--
作者：
Dong, F.;Huang, Y.;Tong, D.
滋养层细胞在胚胎着床和胎盘形成过程中起重要作用。许多有毒化合物可诱导滋养层细胞功能障碍，导致着床失败或早期胚胎丢失。原代滋养层细胞的有限寿命限制了某些毒理学化合物对体外滋养层细胞长期影响的研究。本研究采用密度梯度离心和特异性免疫亲和纯化法对原代山羊滋养层细胞进行纯化。然后，通过转染含有人端粒酶逆转录酶（hTERT）基因的质粒使纯化的细胞永生化。转染hTERT基因的山羊滋养层细胞（hTERT-GTC）可稳定表达hTERT基因，并表现出较高的端粒酶活性，可持续增殖至50代，无衰老迹象。永生化的山羊滋养层细胞仍具有正常原代山羊滋养层细胞的基本和关键特性，表达特异性细胞内标记物细胞角蛋白7（CK-7），分泌绒毛膜促性腺激素β亚基（CG-β）和胎盘催乳素（PL）。进一步研究发现，永生化山羊滋养层细胞表达波形蛋白和非经典MHC I类抗原，并具有侵袭性表型，提示永生化山羊滋养层细胞与人绒毛外滋养层细胞相似。此外，该细胞系在体内或体外均未显示肿瘤转化。结论：hTERT转染的永生化山羊滋养层细胞保留了原代滋养层细胞的基本特征，为研究某些毒物对滋养层细胞的影响提供了一个有用的模型。(C)2013爱思唯尔有限公司保留所有权利。
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 beta-subunit (CG-beta) 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 transfection 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. (C) 2013 Elsevier Ltd. All rights reserved.