[Gonadotropin stimulation of progesterone production of granulosa cells derived from in vitro fertilization procedures: comparison of the in vitro biopotency of recombinant follicle stimulating hormone, recombinant luteinizing hormone nad recombinant human chorionic gonadotropin].
The aim of this study was to investigate the effects of recombinant human gonadotrophins (follicle stimulating hormone [rFSH], luteinizing hormone [rLH] and human chorionic gonadotrophin [rhCG]) on granulosa-lutein cell progesterone production in long-term culture. Cultures were maintained for 9 days including a preincubation period for 3 days. Cells obtained from gonadotrophin-stimulated cycles produced increasing amounts of progesterone during the preincubation period reaching a maximum concentration on day 3. Granulosa cells in absence of serum secreted significantly lower levels of progesterone than in its presence and showed moderate responses to rhCG. Addition of serum (foetal calf serum, FCS) to the culture medium enhanced progesterone output in both control and rhCG-stimulated cultures in a dose-dependent manner. However, in absence of rhCG, granulosa cell progesterone production declined towards the end of culture even in the presence of constantly high FCS levels. All three recombinant gonadotrophins stimulated progesterone accumulation. Recombinant FSH and rLH, applied in the dose interval of 0.001-0.01-0.1 IU/ml, caused clear dose-related increases in progesterone production. Progesterone accumulation was also significantly augmented by the presence of rhCG (range of doses 0.01-0.1-1-10 IU/ml), but this effect was dose-dependent only between 0.1-10 IU/ml dose intervals with the maximum stimulation occurred at the dose of 0.1 IU/ml rhCG. From our results it can be concluded that granulosa cells require both serum supplementation and gonadotrophin stimulation for optimal progesterone synthesis. Recombinant FSH, completely devoid of LH activity, was equivalent to rLH and rhCG in terms of stimulation of progesterone production of luteinized human granulosa cells.
Földesi
I
I
Albert Ludwig Egyetem, Szülészeti és Nógyógyászati Klinika, Freiburg.
Brekwoldt
M
M
Neulen
J
J
D002478:Cells, Cultured; D006063:Chorionic Gonadotropin / Q000494:pharmacology*; D005260:Female; D005307:Fertilization in Vitro; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D006107:Granulosa Cells / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D007986:Luteinizing Hormone / Q000494:pharmacology*; D011374:Progesterone / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000494:pharmacology
D006063:Chorionic Gonadotropin; D011994:Recombinant Proteins; D011374:Progesterone; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone