ANCHORAGE-INDEPENDENT CULTURE OF BOVINE GRANULOSA-CELLS - THE EFFECTS OF BASIC FIBROBLAST GROWTH-FACTOR AND DIBUTYRYL-CAMP ON CELL-DIVISION AND DIFFERENTIATION
ANCHORAGE-INDEPENDENT CULTURE OF BOVINE GRANULOSA-CELLS - THE EFFECTS OF BASIC FIBROBLAST GROWTH-FACTOR AND DIBUTYRYL-CAMP ON CELL-DIVISION AND DIFFERENTIATION
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DOI：
10.1006/excr.1994.1084
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发表时间：
1994-04-01
影响因子：
3.7
通讯作者：
RODGERS, RJ
中科院分区：
文献类型：
--
作者：
LAVRANOS, TC;RODGERS, HF;RODGERS, RJ
在卵巢卵泡发生过程中，颗粒细胞在与基质细胞和其他颗粒细胞接触时分裂。然而，排卵后，它们停止分裂并分化成大黄体细胞。当单层培养时，颗粒细胞自发分化为黄体细胞，从而混淆了颗粒细胞体外滤泡功能（例如细胞分裂）的研究。我们发现牛颗粒细胞能够在由软琼脂和甲基纤维素覆盖层组成的不依赖贴壁的培养系统中分裂。细胞以集落形式生长并保留滤泡颗粒细胞的超微结构特征。它们还分泌具有基底层特征的细胞外基质。颗粒细胞对碱性成纤维细胞生长因子 (bFGF) 的反应呈剂量和时间依赖性，每个培养皿的 DNA 水平增加了三到五倍（50 ng bFGF/ml，持续 14 天）。这种促有丝分裂作用被二丁酰 cAMP (dbcAMP) (1 mM) 抑制。在 dbcAMP 存在下，细胞显着肥大，不分泌细胞外基质，并形成黄体细胞的超微结构特征。他们还分泌三倍以上的黄体酮。该系统提供了研究培养物中颗粒细胞的滤泡功能的能力。 (C) 1994 年学术出版社
During ovarian folliculogenesis granulosa cells divide while in contact with stromal cells and other granulosa cells. Following ovulation, however, they cease dividing and differentiate into large luteal cells. When cultured in monolayer, granulosa cells spontaneously differentiate into luteal cells, thus confounding the study of the follicular functions of granulosa cells in vitro, such as cell division. We have found that bovine granulosa cells were able to divide in an anchorage-independent culture system consisting of soft agar and an overlay of methylcellulose. The cells grew in colonies and retained the ultrastructural features of follicular granulosa cells. They also secreted an extracellular matrix with features of basal lamina. The granulosa cells responded to basic fibroblast growth factor (bFGF) in a dose- and time-dependent manner, with a three- to fivefold increase (at 50 ng bFGF/ml for 14 days) in the level of DNA per dish. This mitogenic effect was inhibited by dibutyryl cAMP (dbcAMP) (1 mM). In the presence of dbcAMP the cells hypertrophied considerably, did not secrete extracellular matrix, and developed the ultrastructural features of luteal cells. They also secreted threefold more progesterone. This system offers the ability to study the follicular functions of granulosa cells in culture. (C) 1994 Academic Press, Inc.