Illumina HiSeq 2000 sequencing; small non-coding RNA sequencing of human cumulus cells from patients with or without Diminished ovarian reserve
small non-coding RNA sequencing of human cumulus cells from patients with or without Diminished ovarian reserve
Cumulus cells were obtained frome two groups of women (DOR group and control group) of comparable age undergoing ovarian hyperstimulation for IVF cycles. A total of 10 women were recruited, and the clinclic characteristics are presented in Table 1. Follicular development was monitored by vaginal ultrasound. All patients were administered human chorionic gonadotropin (10 000 IU Pregnyl, New Jersey; Organon or 0.25 mg Ovitrelle; Serono, Geneva) 34 to 36 hours before transvaginal oocyte collection under ultrasound guidance. Only the cumulusâoocyte complexes with metaphase II (MII) oocytes were included in this study. All patients were of Chinese Han. Cumulus oocyte complexes (COCs) were washed in phosphate-buffered saline and trimmed with a 30-gauge needle. The cumulus oophorus cells were collected and pooled by centrifuging at 15006g for 8 min and immediately frozen in liquid nitrogen until use. All the ten patients were stimulated with the standard long gonadotrophin-releasing hormone agonist (GnRHa, Diphereline; Ipsen Pharma. Biotech, Signes, France) protocol combined with the administration of recombinant FSH (Gonal-f, Merk Serono SA, Geneva, Switzerland). For oocyte retrieve, all patients underwent ovarian puncture (OPU) of follicles larger than 15 mm after 36 h of administration of 10 000 IU human chorionic gonadotrophin (hCG, LiZhu Pharma, ZhuHai, China). Only the cumulus'oocyte complexs with Metaphase II (MII) oocytes were included in this study. Total RNAs were extracted from the human CRCs and COCs using TRIzol reagents (Invitrogen). Library preparation and sequencing was performed at BGI-Shenzhen.
Experimental Factor: diminished ovarian reserve
disease