永生化山羊子宫内膜上皮细胞系的建立及其生物学特性

发布时间：2018-03-19 00:17

本文选题：山羊　切入点：人端粒酶逆转录酶催化亚单位　出处：《中国农业大学学报》2017年02期 　论文类型：期刊论文

【摘要】：为建立永生化山羊子宫内膜上皮细胞系并研究其生物学特性,将人端粒酶逆转录酶催化亚单位(Human telomerase reverse transcrip tase,hTERT)导入原代培养的山羊子宫内膜上皮细胞(Endometrium epithelial cells,EECs),经G418筛选培养,以获得一株永生化山羊子宫内膜上皮细胞(hTERT-EECs);利用流式细胞仪检测其生长周期、凋亡情况及倍体情况;利用MTT法检测其血清依赖性;软琼脂克隆形成试验检测其在半固体培养基中的克隆形成能力;裸鼠致瘤实验检测其成瘤性。结果显示:hTERT-EECs处于S期的细胞(43.2%±2.3%)大于2代EECs(38.7%±1.2%),差异显著(P0.05);hTERT-EECs的凋亡率(1.1%±0.2%)明显低于2代EECs(3.6%±0.5%)(P0.05);倍体分析结果表明hTERT-EECs为二倍体细胞;血清依赖性检测证明hTERT-EECs仍有血清依赖性;软琼脂中无克隆形成能力;对于裸鼠无致瘤性。证明hTERT-EECs具有更强的生长活性,基本属于正常细胞,不具有潜在致瘤性。
[Abstract]:In order to establish an immortalized goat endometrial epithelial cell line and study its biological characteristics, human telomerase reverse transcrip tasehTERT was introduced into the primary cultured goat endometrial epithelial cell line Endometrium epithelial cells EECs, and cultured by G418. In order to obtain an immortalized goat endometrial epithelial cell line hTERT-EECs, the growth cycle, apoptosis and ploidy were detected by flow cytometry, and serum dependence was detected by MTT assay. The clone forming ability of soft Agar was tested in semisolid medium. The results of tumorigenicity test in nude mice showed that the apoptosis rate of hTERT-EECs in S phase (43.2% 卤2.3%) was greater than that of second generation EECs (38.7% 卤1.2%), and the apoptosis rate of hTERT-EECs was significantly lower than that of second generation EECs (3.6% 卤0.5%). Serum dependence test showed that hTERT-EECs still had serum-dependent, no clone forming ability in soft Agar, and no tumorigenicity in nude mice. It was proved that hTERT-EECs had stronger growth activity and belonged to normal cells, and had no potential tumorigenicity.
【作者单位】：西藏农牧学院动物科学学院;
【基金】：西藏自治区自然科学基金(2015212-14-34) 2015年西藏自治区高校临床兽医学重点实验室建设资助项目
【分类号】：S827

【相似文献】