Characterization of an immortalized human granulosa cell line (COV434)
Characterization of an immortalized human granulosa cell line (COV434)
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DOI：
10.1093/molehr/6.2.146
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发表时间：
2000-02-01
影响因子：
4
通讯作者：
De Geyter, C
中科院分区：
文献类型：
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作者：
Zhang, H;Vollmer, M;De Geyter, C
我们研究了一个永生化的颗粒细胞系（COV 434）的生物学特性，它可能被用来研究卵泡和卵母细胞的体外成熟。颗粒细胞功能被定义为由三种不同的特性组成：（i）响应促卵泡激素（FSH）产生17 β-雌二醇;（ii）存在特定的细胞凋亡分子标记物，能够诱导卵泡闭锁;（iii）与卵母细胞周围细胞形成细胞间连接的能力。向补充有10%胎牛血清和4-雄烯-3，17-二酮的培养基中添加FSH导致COV 434颗粒细胞增殖和17 β-雌二醇合成增加，表明这些细胞中存在FSH受体和细胞色素P450芳香酶。未检测到促黄体生成素（LH）受体。在COV 434颗粒细胞和用促性腺激素刺激体外受精的患者的颗粒细胞中发现了各种凋亡相关基因的相似表达，从而表明永生化的COV 434颗粒细胞能够维持凋亡。在COV 434颗粒细胞与含有未成熟卵母细胞的卵丘细胞共培养期间形成多个细胞间连接，但不与缺乏卵母细胞的卵丘细胞共培养。用扫描电子显微镜和共聚焦光学显微镜对细胞间连接进行了详细的形态学分析，证明了细长结构的存在。结论：永生化人卵巢颗粒细胞系COV 434可用于卵泡发育的实验研究。
We have investigated the biological characteristics of an immortalized granulosa cell line (COV434), which may be used to study follicular and oocyte maturation in vitro. Granulosa cell function was defined as consisting of three distinct properties: (i) production of 17 beta-oestradiol in response to follicle stimulating hormone (FSH); (ii) presence of specific molecular markers of apoptosis enabling the induction of follicular atresia; and (iii) capacity to form intercellular connections with cells surrounding an oocyte. The addition of FSH to the culture medium supplemented with 10% fetal calf serum and 4-androstene-3,17-dione resulted in proliferation of the COV434 granulosa cells and in an increased synthesis of 17 beta-oestradiol, indicating the presence of the FSH receptor and cytochrome P450 aromatase in these cells. The receptor for luteinizing hormone (LH) was undetectable. Similar expression of various apoptosis-associated genes was found in COV434 granulosa cells and in granulosa cells of patients stimulated with gonadotrophins for in-vitro fertilization, thus indicating that the immortalized COV434 granulosa cells were able to sustain apoptosis. Multiple intercellular connections were formed during co-culture of COV434 granulosa cells with cumulus cells containing an immature oocyte but not with cumulus cells devoid of an oocyte. Detailed morphological analysis of the intercellular connections with scanning electron microscopy and confocal light microscopy demonstrated the presence of long slender structures. It is concluded that the immortalized human granulosa cell line COV434 may be useful for experimental studies on follicular development.