Abstract
Purpose
The in-vitro environment influences oocyte competence and gene expression in cumulus cells and oocytes. Effects of culturing under non-attachment conditions and varying follicle exposure to FSH were investigated at the mRNA level and on oocyte developmental capacity.
Methods
Quantitative PCR analysis of Gdf9, Mater, Nmp2 (in oocytes), Lhcgr and Amh (in cumulus cells), and oocyte developmental competence after in vitro follicle culture were evaluated.
Results
Follicle survival (98.7%) and polar body rate (94%) were similar for all conditions. Estradiol and progesterone production were significantly lower in non-attachment follicles (10-fold and 3-fold, respectively). Under non-attachment conditions, a higher two-cell rate (69.9%) and total blastocyst yield (48.5%) were obtained and, by decreasing FSH levels during culture, Lhcgr transcripts were significantly reduced to levels similar to in-vivo. Levels of oocyte-specific transcripts were not significantly influenced by in-vitro conditions.
Conclusion
Non-attachment conditions influence follicle steroid secretory capacity and, together with dynamic FSH doses, positively influence cumulus cell gene expression and oocyte developmental competence.
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Acknowledgements
The authors wish to acknowledge Mrs. Katy Billooye for her valuable technical assistance with staining for alkaline phosphatase in theca cells and for performing the radioimmunoassays on the conditioned media. The authors are very grateful to Ms. Sandra De Schaepdryver for her editorial support.
The authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported.
Financial support
This work was supported by The Belgian Foundation Against Cancer (project no. 221.2008).
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Flor Sánchez and Sergio Romero contributed equally to this work.
Capsule
Non-attached follicle culture and decreased FSH levels modulate follicle steroidogenesis and positively influence gene expression in cumulus cells and oocyte developmental capacity.
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Sánchez, F., Romero, S., Albuz, F.K. et al. In vitro follicle growth under non-attachment conditions and decreased FSH levels reduces Lhcgr expression in cumulus cells and promotes oocyte developmental competence. J Assist Reprod Genet 29, 141–152 (2012). https://doi.org/10.1007/s10815-011-9690-x
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DOI: https://doi.org/10.1007/s10815-011-9690-x