Effect of apoptosis and reactive oxygen species production in human granulosa cells on oocyte fertilization and blastocyst development.
OBJECTIVE
The aim was to establish the impact of human granulosa cell apoptosis and reactive oxygen species (ROS) production on fertilization competence of the oocyte, embryo developmental stage and implantation rate.


METHODS
Thirty women undergoing IVF-ET for tubal factor infertility were included; GnRH antagonists and gonadotrophins were used for ovarian stimulation. Granulosa cells were isolated from each aspirated follicle using gradient centrifugation. Apoptosis was studied by flow cytometry using annexin V and propidium iodide. ROS production was studied with hydroethidine staining and analyzed by flow cytometry.


RESULTS
There were no differences in characteristics of granulosa cells between the follicles with fertilized and non-fertilized oocytes. The analyzed characteristics of granulosa cells in corresponding follicles had no effect on embryo developmental stage on day 5. The percentage of ROS producing granulosa cells was lower in the follicles giving rise to blastocysts that resulted in implantation compared to those that did not (39.9% versus 69.9%, P = 0.031).


CONCLUSIONS
Apoptosis and ROS production in granulosa cells have no significant impact on fertilization and do not correlate with the development of blastocysts. An increased percentage of ROS producing granulosa cells results in fewer oocytes retrieved and diminishes implantation rate.
Jancar
Nina
N
Reproductive Unit, Department of Obstetrics and Gynecology, University Medical Centre Ljubljana, Slajmerjeva 3, SI-1000 Ljubljana, Slovenia. nina.jancar@kclj.si
Kopitar
Andreja N
AN
Ihan
Alojz
A
Virant Klun
Irma
I
Bokal
Eda Vrtacnik
EV
D000328:Adult; D017209:Apoptosis; D001755:Blastocyst / Q000502:physiology*; D047108:Embryonic Development / Q000502:physiology*; D005260:Female; D005306:Fertilization / Q000502:physiology*; D005307:Fertilization in Vitro; D006107:Granulosa Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D007247:Infertility, Female / Q000628:therapy; D009865:Oocytes / Q000254:growth & development*; D017382:Reactive Oxygen Species / Q000378:metabolism*
D017382:Reactive Oxygen Species