Archives of Gynecology and Obstetrics

, Volume 270, Issue 3, pp 174–178 | Cite as

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

  • Jochen ReinsbergEmail author
  • Dietlind Ackermann
  • Hans van der Ven
Original Article



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.


Absorption Assisted reproduction Estradiol Human Tissue culture 


  1. 1.
    Augustin-Voss HG, Pauli BU (1992) Quantitative analysis of autocrine-regulated, matrix-induced, and tumor cell-stimulated endothelial cell migration using a silicone template compartmentalization technique. Exp Cell Res 198:221–228PubMedGoogle Scholar
  2. 2.
    Bakker D, van Blitterswijk CA, Daems WT, Grote JJ (1988) Biocompatibility of six elastomers in vitro. Biomed Mater Res 22:423–439Google Scholar
  3. 3.
    Balmaceda JP, Pool TB, Arana JB, Heitman TS, Asch RH (1984) Successful in vitro fertilization and embryo transfer in cynomolgus monkeys. Fertil Steril 42:791–795PubMedGoogle Scholar
  4. 4.
    Berg DK, Thompson JG, Asher GW (2002) Development of in vitro embryo production system for red deer (Cervus elaphus). II. The timing of in vitro nuclear oocyte maturation. Anim Reprod Sci 70:77–84CrossRefPubMedGoogle Scholar
  5. 5.
    Cekleniak NA, Combelles CMH, Ganz DA, Fung J, Albertini DF, Racowsky C (2001) A novel system for in vitro maturation of human oocytes. Fertil Steril 75:1185–1193CrossRefPubMedGoogle Scholar
  6. 6.
    Chauvel-Lebret DJ, Pellen-Mussi P, Auroy P, Bonnaure-Mallet M (1999) Evaluation of the in vitro biocompatibility of various elastomers. Biomaterials 20:291–299CrossRefPubMedGoogle Scholar
  7. 7.
    Cortvrindt R, Smitz J, Van Steirteghem AC (1996) In-vitro maturation, fertilization and embryo development of immature oocytes from early preantral follicles from prepubertal mice in a simplified culture system. Hum Reprod 11:2656–2666PubMedGoogle Scholar
  8. 8.
    De Vantery Arrighi C, Lucas H, El-Mowafi D, Campana A, Chardonnes D (2001) Effects of human hydrosalpinx fluid on in-vitro murine fertilization. Hum Reprod 16:676–682CrossRefPubMedGoogle Scholar
  9. 9.
    Englund DE, Victor A, Johansson ED (1981) Pharmacokinetics and pharmacodynamic effects of vaginal oestradiol administration from silastic rings in post-menopausal women. Maturitas 3:125–133PubMedGoogle Scholar
  10. 10.
    Goud PT, Goud AP, Qian C, Laverge H, Van der Elst J, De Sutter P, Dhont M (1998) In-vitro maturation of human germinal vesicle stage oocytes: role of cumulus cells and epidermal growth factor in the culture medium. Hum Reprod 13:1638–1644CrossRefPubMedGoogle Scholar
  11. 11.
    Grondahl C, Hansen TH, Marky-Nielsen K, Ottesen JL, Hyttel P (2000) Human oocyte maturation in vitro is stimulated by meiosis-activating sterol. Hum Reprod 15:3–10CrossRefGoogle Scholar
  12. 12.
    Hofmann J, Raether W (1990) Improved techniques for the in vitro cultivation of Eimeria tenella in primary chick kidney cells. Parasitol Res 76:479–486PubMedGoogle Scholar
  13. 13.
    Hwu Y, Lee RK, Chen C, Su J, Chen Y, Lin S (1998) Development of hatching blastocysts from immature human oocytes following in-vitro maturation and fertilization using a co-culture system. Hum Reprod 13:1916–1921CrossRefPubMedGoogle Scholar
  14. 14.
    Koos RD, Feiertag MA (1989) The effect of reduced oxygen tension on progesterone accumulation in rat granulosa cell cultures. Steroids 54:553–562CrossRefPubMedGoogle Scholar
  15. 15.
    Liu LPS, Chan STH, Ho PC, Yeung WSB (1998) Partial purification of embryotrophic factors from human oviductal cells. Hum Reprod 13:1613–1619CrossRefPubMedGoogle Scholar
  16. 16.
    Miller KF, Pursel VG (1987) Absorption of compounds in medium by the oil covering microdrop cultures. Gamete Res 17:57–61PubMedGoogle Scholar
  17. 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
  18. 18.
    Milligan SR, Cohen PE (1994) Silastic implants for delivering physiological concentrations of progesterone to mice. Reprod Fertil Dev 6:235–239PubMedGoogle Scholar
  19. 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
  20. 20.
    Peluso JJ, Pappalardo A (1994) Progesterone and cell-cell adhesion interact to regulate rat granulosa cell apoptosis. Biochem Cell Biol 72:547–551PubMedGoogle Scholar
  21. 21.
    Seelbach A, Fissler B, Madle S (1993) Further evaluation of a modified micronucleus assay with V79 cells for detection of aneugenic effects. Mutat Res 303:163–169PubMedGoogle Scholar
  22. 22.
    Smitz J, Cortvrindt R, van Steirteghem AC (1996) Normal oxygen atmosphere is essential for the solitary long-term culture of early preantral mouse follicles. Mol Reprod Dev 45:466–475CrossRefPubMedGoogle Scholar
  23. 23.
    Speit G, Haupter S, Pentz S (1990) An improved procedure for the cultivation and in situ chromosome preparation of monolayer cell cultures. Cytobios 62:39–44PubMedGoogle Scholar
  24. 24.
    Wrenzycki C, Herrmann D, Keskintepe L, Martins A, Sirisathien S, Brackett B, Niemann H (2001) Effects of culture system and protein supplementation on mRNA expression in pre-implantation bovine embryos. Hum Reprod 16:893–901CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • Jochen Reinsberg
    • 1
    Email author
  • Dietlind Ackermann
    • 1
  • Hans van der Ven
    • 1
  1. 1.Department of Gynecological Endocrinology and Reproductive MedicineUniversity of BonnBonnGermany

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