title: Pitfalls in assessment of progesterone production by granulosa cells cultured in contact with silicone rubber or paraffin oil.
abstract: 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.
authors.affiliation: Department of Gynecological Endocrinology and Reproductive Medicine, University of Bonn, Sigmund-Freud-Strasse 25, 53127 Bonn, Germany. Jochen.Reinsberg@ukb.uni-bonn.de
authors.forename: Jochen
authors.initials: J
authors.lastname: Reinsberg
authors.forename: Dietlind
authors.initials: D
authors.lastname: Ackermann
authors.forename: Hans
authors.initials: H
authors.lastname: van der Ven
mesh_terms: 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
chemical_list: D009821:Oils; D012826:Silicone Elastomers; D011374:Progesterone; D004958:Estradiol; D010232:Paraffin; C015418:paraffin oils