Reinsberg Jochen, Ackermann Dietlind, van der Ven Hans
Department of Gynecological Endocrinology and Reproductive Medicine, University of Bonn, Sigmund-Freud-Strasse 25, 53127 Bonn, Germany.
Arch Gynecol Obstet. 2004 Nov;270(3):174-8. doi: 10.1007/s00404-003-0539-0. Epub 2003 Aug 19.
The influence of silicone templates, used for compartmentalization of culture dishes, on progesterone accumulation in granulosa cell cultures is studied and compared with the effect of paraffin oil, which is frequently used to cover oocyte/embryo cultures.
Human granulosa-lutein cells were cultured in culture dishes compartmentalized by silicone templates, or in polystyrene plates under paraffin oil. Progesterone concentrations in the culture supernatant were compared with controls cultured in polystyrene plates.
The progesterone concentration in culture supernatant was grossly reduced in silicone template cultures (2+/-0.7% of control). No inhibitory activity was identified in medium conditioned by preincubation with silicone rubber, but progesterone was absorbed from spiked medium incubated in silicone templates (recovery <2%). Progesterone concentration in culture supernatant was also reduced by a paraffin oil overlay (38+/-3% of control). From steroid spiked microdrops under oil, <2% of progesterone and 85+/-4% of estradiol was recovered.
The steroidogenesis of cells cultured in silicone templates or under oil cannot be assessed correctly. It has to be considered that the concentration of lipophilic compounds may be grossly changed due to absorption by silicone rubber or paraffin oil.
研究用于培养皿分隔的硅胶模板对颗粒细胞培养中孕酮积累的影响，并与常用于覆盖卵母细胞/胚胎培养的石蜡油的作用进行比较。
将人颗粒黄体细胞培养于用硅胶模板分隔的培养皿中，或在石蜡油覆盖下培养于聚苯乙烯板中。将培养上清液中的孕酮浓度与培养于聚苯乙烯板中的对照进行比较。
硅胶模板培养中培养上清液中的孕酮浓度大幅降低（为对照的2±0.7%）。在用硅橡胶预孵育的条件培养基中未发现抑制活性，但孕酮从在硅胶模板中孵育的加标培养基中被吸收（回收率<2%）。石蜡油覆盖也降低了培养上清液中的孕酮浓度（为对照的38±3%）。从油下的类固醇加标微滴中，孕酮回收率<2%，雌二醇回收率为85±4%。
无法正确评估在硅胶模板中或油下培养的细胞的类固醇生成。必须考虑到亲脂性化合物的浓度可能因被硅橡胶或石蜡油吸收而大幅改变。
