Reference : In vitro evaluation of the anti-apoptotic drug Z-VAD-FMK on human ovarian granulosa c...
Scientific journals : Article
Human health sciences : Reproductive medicine (gynecology, andrology, obstetrics)
http://hdl.handle.net/2268/186693
In vitro evaluation of the anti-apoptotic drug Z-VAD-FMK on human ovarian granulosa cell lines for further use in ovarian tissue transplantation.
English
Fransolet, Maïté mailto [Université de Liège > Département des sciences cliniques > Gynécologie - Obstétrique >]
HENRY, Laurie mailto [Centre Hospitalier Universitaire de Liège - CHU > > Gynécologie-Obstétrique CHR >]
Labied, Soraya [> >]
Noël, Agnès mailto [Université de Liège > Département des sciences cliniques > Labo de biologie des tumeurs et du développement >]
Nisolle, Michelle mailto [Université de Liège > Département des sciences cliniques > Gynécologie - Obstétrique >]
Munaut, Carine mailto [Université de Liège > Département des sciences cliniques > Labo de biologie des tumeurs et du développement >]
2015
Journal of Assisted Reproduction and Genetics
Kluwer Academic/Plenum Publishers
Yes (verified by ORBi)
International
1058-0468
1573-7330
New York
NY
[en] Ovarian granulosa cells ; Z-VAD-FMK ; Etoposide ; Hypoxia; ; Ovarian transplantation ; Fertility preservation
[en] PURPOSE: Because ovarian granulosa cells are essential for oocyte survival, we examined three human granulosa cell lines as models to evaluate the ability of the pan-caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (Z-VAD-FMK) to prevent primordial follicle loss after ovarian tissue transplantation. METHODS: To validate the efficacy of Z-VAD-FMK, three human granulosa cell lines (GC1a, HGL5, COV434) were treated for 48 h with etoposide (50 mug/ml) and/or Z-VAD-FMK (50 muM) under normoxic conditions. To mimic the ischemic phase that occurs after ovarian fragment transplantation, cells were cultured without serum under hypoxia (1 % O2) and treated with Z-VAD-FMK. The metabolic activity of the cells was evaluated by WST-1 assay. Cell viability was determined by FACS analyses. The expression of apoptosis-related molecules was assessed by RT-qPCR and Western blot analyses. RESULTS: Our assessment of metabolic activity and FACS analyses in the normoxic experiments indicate that Z-VAD-FMK protects granulosa cells from etoposide-induced cell death. When cells are exposed to hypoxia and serum starvation, their metabolic activity is reduced. However, Z-VAD-FMK does not provide a protective effect. In the hypoxic experiments, the number of viable cells was not modulated, and we did not observe any modifications in the expressions of apoptosis-related molecules (p53, Bax, Bcl-xl, and poly (ADP-ribose) polymerase (PARP)). CONCLUSION: The death of granulosa cell lines was not induced in our ischemic model. Therefore, a protective effect of Z-VAD-FMK in vitro for further use in ovarian tissue transplantation could not be directly confirmed. It will be of interest to potentially use Z-VAD-FMK in vivo in xenograft models.
http://hdl.handle.net/2268/186693
10.1007/s10815-015-0536-9

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