Establishment and characterization of an immortalized human oviductal cell line
Establishment and characterization of an immortalized human oviductal cell line
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DOI：
10.1002/mrd.1046
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发表时间：
2001-08-01
影响因子：
2.5
通讯作者：
Yeung, WSB
中科院分区：
文献类型：
--
作者：
Lee, YL;Lee, KF;Yeung, WSB
人输卵管细胞刺激胚胎体外发育部分是通过产生胚胎营养糖蛋白。这些糖蛋白的身份尚不清楚，主要是因为输卵管样品有限，培养的亲本输卵管细胞不能产生足够量的胚胎营养因子进行表征。本研究通过逆转录病毒表达，将人乳头瘤病毒16型E6/E7开放阅读框（ORF）导入人输卵管上皮细胞（OE），使其永生化。该细胞系（OE-E6/E7）的特征与亲本细胞相似，但在OE-E6/E7中未发现HPV 16 E6/E7 DNA，且在两种细胞中均发现人输卵管特异性糖蛋白、雌激素受体和细胞角蛋白。OE和OE-E6/E7均具有端粒酶活性，但前者活性较低。结果表明，OE-E6/E7共培养的小鼠早期胚胎的囊胚形成率明显高于单独培养的小鼠早期胚胎，且OE-E6/E7共培养的小鼠早期胚胎的囊胚形成率明显高于单独培养的小鼠早期胚胎（52%对32%）和囊胚直径（113.0 +/- 2.07 μ m对83.9 +/- 5.23 μ m）。该永生化细胞系可作为一个连续、稳定的体外系统用于输卵管胚胎营养活性的研究。(C)2001 Wiley-Liss，Inc.
Human oviductal cells stimulate embryo development in vitro partly by the production of embryotrophic glycoproteins. The identity of these glycoproteins is not yet known mainly because oviductal samples are limited and that the cultured parental oviductal cells cannot produce sufficient amount of embryotrophic factors for characterization. In this study, human oviductal epithelial cells (OE) were immortalized by HPV 16 E6/E7 open reading frame (ORF) by retroviral expression. The characteristics of this immortalized cell line (OE-E6/E7) were com pared to the parental OE. H PV 16 E6/E7 DNA was found only in OE-E6/E7 but not in OE. Human oviduct-specific glycoprotein, estrogen receptors, and cytokeratin were found in both cell types. Both OE and OE-E6/E7 possessed telomerase activities but the former had much lower activity. OE-E6/E7 also produced glycoproteins with chromatographic behavior similar to the embryotrophic glycoproteins derived from OE. These results showed that OE-E6/E7 retained a number of characteristics of OE. The development of preimplantation mouse embryo was significantly better after coculture with OE-E6/E7 when compared to medium alone culture in term of blastulation rates (52% vs. 32%) and blastocyst diameter (113.0 +/- 2.07 mum vs. 83.9 +/- 5.23 mum). This immortalized cell line can be used as a continuous and stable in vitro system for the study of the oviductal embryotrophic activity. (C) 2001 Wiley-Liss, Inc.