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  • The transcriptome of corona radiata cells from individual MІІ oocytes that after ICSI developed to embryos selected for transfer: PCOS women compared to healthy women.

The transcriptome of corona radiata cells from individual MІІ oocytes that after ICSI developed to embryos selected for transfer: PCOS women compared to healthy women.

Journal of ovarian research (2014-11-30)
Marie Louise Wissing, Si Brask Sonne, David Westergaard, Kho do Nguyen, Kirstine Belling, Thomas Høst, Anne Lis Mikkelsen
ABSTRACT

Corona radiata cells (CRCs) refer to the fraction of cumulus cells just adjacent to the oocyte. The CRCs are closely connected to the oocyte throughout maturation and their gene expression profiles might reflect oocyte quality. Polycystic ovary syndrome (PCOS) is a common cause of infertility. It is controversial whether PCOS associate with diminished oocyte quality. The purpose of this study was to compare individual human CRC samples between PCOS patients and controls. All patients were stimulated by the long gonadotropin-releasing hormone (GnRH) agonist protocol. The CRC samples originated from individual oocytes developing into embryos selected for transfer. CRCs were isolated in a two-step denudation procedure, separating outer cumulus cells from the inner CRCs. Extracted RNA was amplified and transcriptome profiling was performed with Human Agilent® arrays. The transcriptomes of CRCs showed no individual genes with significant differential expression between PCOS and controls, but gene set enrichment analysis identified several cell cycle- and DNA replication pathways overexpressed in PCOS CRCs (FDR < 0.05). Five of the genes contributing to the up-regulated cell cycle pathways in the PCOS CRCs were selected for qRT-PCR validation in ten PCOS and ten control CRC samples. qRT-PCR confirmed significant up-regulation in PCOS CRCs of cell cycle progression genes HIST1H4C (FC = 2.7), UBE2C (FC = 2.6) and cell cycle related transcription factor E2F4 (FC = 2.5). The overexpression of cell cycle-related genes and cell cycle pathways in PCOS CRCs could indicate a disturbed or delayed final maturation and differentiation of the CRCs in response to the human chorionic gonadotropin (hCG) surge. However, this had no effect on the in vitro development of the corresponding embryos. Future studies are needed to clarify whether the up-regulated cell cycle pathways in PCOS CRCs have any clinical implications.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
17α-Ethynylestradiol, ≥98%
Ethinylestradiol, European Pharmacopoeia (EP) Reference Standard
Supelco
Ethinyl Estradiol, Pharmaceutical Secondary Standard; Certified Reference Material
Ethinylestradiol, British Pharmacopoeia (BP) Assay Standard
USP
Ethinyl estradiol, United States Pharmacopeia (USP) Reference Standard
Desogestrel, European Pharmacopoeia (EP) Reference Standard
Desogestrel for system suitability, European Pharmacopoeia (EP) Reference Standard
Ethinylestradiol for system suitability, European Pharmacopoeia (EP) Reference Standard