Apoptosis of cultured granulosa-lutein cells is reduced by insulin-like growth factor I and may correlate with embryo fragmentation and pregnancy rate.
OBJECTIVE
To correlate apoptosis of cultured human granulosa-lutein cells (GL cells) with the outcome of IVF (embryo fragmentation and pregnancy rate) and to study the effect of insulin and insulin-like growth factor I (IGF-I) on apoptosis.


METHODS
In vitro assays.


METHODS
University laboratory and private IVF center.


METHODS
Eighty-one women undergoing IVF.


METHODS
Purified human GL cells from pooled follicles were cultured for 48 hours in serum-free media with or without insulin and IGF-I. Cumulus cells and mural GL cells were evaluated separately.


METHODS
Detection of apoptosis by using caspACE FITC-VAD-FMK, a fluorescent in situ marker for activated caspases; embryo fragmentation; and pregnancy.


RESULTS
Age younger than 38 years and successful pregnancy were associated with less apoptosis (33.0% +/- 17.2% vs. 43.2% +/- 18.0% and 30.2% +/- 14.0% vs. 40.4% +/- 19.5%, respectively). There was a linear correlation between embryo fragmentation and GL cell apoptosis. Insulin-like growth factor I decreased apoptosis in a dose-dependent fashion. A statistically significant effect (17% decrease) was reached at a dose of 10 nM. Insulin (10 nM) caused a small (8%) decrease in apoptosis, but this effect did not reach statistical significance. Cumulus cells consistently had <3% apoptosis.


CONCLUSIONS
[1] Apoptosis of cultured GL cells may be associated with IVF outcome and ovarian reserve and [2] IGF-I decreases apoptosis of cultured GL cells.
Bencomo
Eija
E
Departamento de Bioquímica y Biología Molecular, Universidad de la Laguna, La Laguna, Tenerife, Spain.
Pérez
Rosa
R
Arteaga
Mari-Francis
MF
Acosta
Elisa
E
Peña
Oscar
O
Lopez
Lourdes
L
Avila
Julio
J
Palumbo
Angela
A
D000328:Adult; D017209:Apoptosis / Q000187:drug effects*; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D004622:Embryo, Mammalian / Q000502:physiology*; D005260:Female; D005307:Fertilization in Vitro; D006801:Humans; D007246:Infertility / Q000503:physiopathology* / Q000628:therapy*; D007328:Insulin / Q000494:pharmacology; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology*; D008184:Luteal Cells / Q000187:drug effects*; D008297:Male; D011247:Pregnancy; D018873:Pregnancy Rate; D016896:Treatment Outcome
D016895:Culture Media, Serum-Free; D007328:Insulin; D007334:Insulin-Like Growth Factor I