INVITRO DIFFERENTIATION OF BOVINE THECA AND GRANULOSA-CELLS INTO SMALL AND LARGE LUTEAL-LIKE CELLS - MORPHOLOGICAL AND FUNCTIONAL-CHARACTERISTICS
INVITRO DIFFERENTIATION OF BOVINE THECA AND GRANULOSA-CELLS INTO SMALL AND LARGE LUTEAL-LIKE CELLS - MORPHOLOGICAL AND FUNCTIONAL-CHARACTERISTICS
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DOI：
10.1095/biolreprod43.6.913
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发表时间：
1990-12-01
影响因子：
3.6
通讯作者：
ABERDAM, E
中科院分区：
文献类型：
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作者：
MEIDAN, R;GIRSH, E;ABERDAM, E
本研究旨在探讨牛卵泡颗粒细胞和卵泡内膜细胞能否在体外被促黄体化为黄体样细胞。在毛喉素（10 μ M）、胰岛素（2 μ g/ml）、胰岛素样生长因子I（100 ng/ml）或这些试剂的组合存在下培养颗粒细胞和卵泡膜细胞9天。在培养的第一天，颗粒细胞和卵泡膜细胞分泌雌二醇和雄烯二酮，分别;孕酮上升后，仅在培养3-5天，并在培养的第九天达到最大值。在毛喉素加胰岛素存在下孵育的细胞表现出从黄体分离的黄体细胞的形态和功能特征。结果发现，两种细胞类型的细胞直径，基础和刺激的前列腺素分泌，和细胞复制的模式是可比的黄体细胞。大量的脂滴和增强的线粒体肾上腺素还表明活跃的类固醇激素合成的黄素化细胞染色。培养9天后，将刺激物撤出，并在基础培养基中再培养5天;黄体化颗粒细胞（LGC）维持孕酮升高，而相反，在黄体化卵泡膜细胞（LTC）中观察到孕酮产生急剧下降。在第9天，用LH（10 ng/ml）、毛喉素（10 μ M）或霍乱毒素（100 ng/ml）攻击细胞3小时，导致4倍;相同的处理不能刺激LGC中的孕酮。这些结果表明，LGC和LCT中类固醇生成的调节似乎是不同的：LTC高度依赖于cAMP升高剂并对cAMP升高剂有反应。另一方面，LGC不能被刺激，但仍然能够在基础条件下维持类固醇生成。
This study was undertaken to investigate whether bovine granulosa and theca interna cells could be luteinized in vitro into luteal-like cells. Granulosa and theca cells were cultured for 9 days in the presence of forskolin (10 .mu.M), insulin (2 .mu.g/ml), insulin-like growth factor I (100 ng/ml), or a combination of these agents. During the first days of culture, granulosa and theca cells secreted estradiol and androstenedione, respectively; progesterone rose only after 3-5 days in culture and reached a maximum on the ninth day of culture. Cells incubated in the presence of forskolin plus insulin exhibited morphological and functional characteristics of luteal cells isolated from the corpus luteum. It was found that cell diameter, basal and stimulated porgesterone secretion, and pattern of cell replication for both cell types were comparable to those of luteal cells. Numerous lipid droplets and intensified mitochondrial adrenodoxin staining also indicated active steroidogenesis in luteinized cells. After 9 days in culture, stimulants were withdrawan, and the culture proceeded in basal medium for an additional 5 days; elevated progesterone that were maintained by luteinized granulosa cells (LGC), whereas in contrast a dramatic drop in progesterone production was observed in luteinized theca cells (LTC). On Day 9, cells were challenged for 3 h with LH (10 ng/ml), forskolin (10 .mu.M), or cholera toxin (100 ng/ml), resulting in a 4-fold; the same treatments failed to stimulate progesterone in LGC. These results demonstrate the regulation demonstrate that regulation of steroidogenesis in LGC and LCT appears to be different: LTC are highly dependent on and responsive to cAMP elevating agents. LGC, on the other hand, are unable to be stimulated but are nonetheless able to sustain steroidogenesis under basal conditions.