Pitfalls in assessment of progesterone production by granulosa cells cultured in contact with silicone rubber or paraffin oil
- 70 Downloads
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.
KeywordsAbsorption Assisted reproduction Estradiol Human Tissue culture
- 2.Bakker D, van Blitterswijk CA, Daems WT, Grote JJ (1988) Biocompatibility of six elastomers in vitro. Biomed Mater Res 22:423–439Google Scholar
- 17.Miller KF, Goldberg JM, Collins RL (1994) Covering embryo cultures with mineral oil alters embryo growth by acting as a sink for an embryotoxic substance. Assist Reprod Genet 11:342–345Google Scholar
- 19.Mussche S, D'Herde K (2001) Contribution of progesterone, follicle stimulating hormone and glucocorticoids in survival of serum-free cultured granulosa cell explants. Endocrinology 169:321–331Google Scholar