Gregoraszczuk E L
Department of Animal Physiology, Jagiellonian University, Krakow, Poland.
Cytotechnology. 1990 Sep;4(2):195-200. doi: 10.1007/BF00365101.
Ovarian cells such as theca interna, granulosa and corpus luteum cells were isolated from pig ovaries and cultured in Erlenmayer flasks (25 ml) containing 3.5 ml of culture medium. The media were replaced every second day and frozen to -20 degrees C for later steroid analysis. The reaggregation of cells was completed within 2-3 days and this was then followed by a period of cell migration. During the subsequent 5-6 day period the reaggregates became larger. The best results were obtained in cultures of isolated theca alone and in combination with granulosa cells, as well as of early corpus luteum cells. Granulosa cells did not aggregate as easily or as completely as the corpus luteum cells. All types of cells investigated were able to secrete progesterone into the culture medium. They secreted more progesterone and for a longer time than cells cultured as monolayers. The aggregate culture seems to be a good model to study the secretion of ovarian cells, by creating the tri-dimensional, and thus more physiological, culture system.
从猪卵巢中分离出卵泡内膜、颗粒细胞和黄体细胞等卵巢细胞,并在装有3.5毫升培养基的三角烧瓶(25毫升)中培养。每隔一天更换培养基,并冷冻至-20摄氏度以备后续的类固醇分析。细胞重聚集在2-3天内完成,随后是一段细胞迁移期。在随后的5-6天内,重聚集体变得更大。单独培养分离出的卵泡内膜细胞以及与颗粒细胞联合培养,还有早期黄体细胞培养,均取得了最佳结果。颗粒细胞不像黄体细胞那样容易聚集或完全聚集。所研究的所有类型细胞都能够将孕酮分泌到培养基中。它们分泌的孕酮比单层培养的细胞更多,且分泌时间更长。聚集培养似乎是通过创建三维且因此更具生理性的培养系统来研究卵巢细胞分泌的良好模型。