Immortalization of goat endometrial epithelial cells by telomerase reverse transcriptase transfection.
Immortalization of goat endometrial epithelial cells by telomerase reverse transcriptase transfection.
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发表时间：
2010
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通讯作者：
Wu Qing-xia;Wang Aihua;Si YongJia;Zhou Limei;Sheng HongXia;Jin Ya-ping
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作者：
Wu Qing-xia;Wang Aihua;Si YongJia;Zhou Limei;Sheng HongXia;Jin Ya-ping
为获得大量具有高度同源性和正常表型功能的山羊子宫内膜上皮细胞，将人端粒酶逆转录酶(HTERT)载体pCI-neo-hTERT导入山羊子宫内膜上皮细胞，筛选并扩增G418抗性克隆，检测稳定细胞克隆的表型、增殖和生长特性。结果显示，转染组细胞融合单层呈鹅卵石样，细胞角蛋白呈阳性反应，免疫细胞化学染色显示细胞端粒酶活性高于未转染组，传代至50代，培养稳定，100nmol/L雌二醇可刺激转染组EEC增殖；50,100nmol/L P4可抑制EEC的增殖。研究表明，端粒酶活性在转染人端粒酶逆转录酶后能被激活，从而获得高同源性和正常表型的细胞。
To obtain a great quantity of goat endometrium epithelial cells(EECs) with highly identity and normal phenotype function,plasmid pCI-neo-hTERT,the human telomerase reverse transcriptase (hTERT) was transfected into goat EECs.Clones resisted G418 were selected and amplified.The phenotype,proliferation and growth characteristic of stably cell clones were detected,also the effect of sex steroids on the proliferation were determined.The results showed that the confluent monolayer of transfected cells looked like cobblestones with contact inhibition.The transfected cells were positive for cytokeratin and had higher telomerase activity than that of untransfected cells by immunocytochemical staining method.It was cultured up to 50 passages and steadily cultured.100 nmol/L E2 can stimulate the proliferation of transfected EEC;50,100 nmol/L P4 can inhibit the proliferation of transfected EEC.The research indicated that telomerase activity can be activated in goat EEC after hTERT transfected,thus the cells with high identity and normal phenotype were obtained.