Pitfalls in assessment of progesterone production by granulosa cells cultured in contact with silicone rubber or paraffin oil.
BACKGROUND
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.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Reinsberg
Jochen
J
Department of Gynecological Endocrinology and Reproductive Medicine, University of Bonn, Sigmund-Freud-Strasse 25, 53127 Bonn, Germany. Jochen.Reinsberg@ukb.uni-bonn.de
Ackermann
Dietlind
D
van der Ven
Hans
H
D018929:Cell Culture Techniques / Q000295:instrumentation*; D002478:Cells, Cultured; D004958:Estradiol / Q000378:metabolism; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods*; D006107:Granulosa Cells / Q000378:metabolism*; D006801:Humans; D009821:Oils; D010232:Paraffin; D011374:Progesterone / Q000378:metabolism*; D012826:Silicone Elastomers
D009821:Oils; D012826:Silicone Elastomers; D011374:Progesterone; D004958:Estradiol; D010232:Paraffin; C015418:paraffin oils