P-281 Single-cell RNA sequencing analysis of granulosa cells in preovulatory follicles in normal ovarian reserve patients undergoing progestin-primed ovarian stimulation and GnRH-antagonist

Abstract Study question Could the difference in controlled ovarian stimulation protocol between progestin-primed and GnRH-antagonist induce change expression genes of human granulosa cells? Summary answer The mitochondrial DNA (mtDNA) gene cells patients who underwent was significantly higher than GnRH-antagonist. What is known already (PPOS) has attracted attention many studies have reported similar pregnancy outcomes for both PPOS (GnRH-ant). However, our previous study demonstrated that rate lower compared to GnRH-ant (38th Hybrid Annual Meeting ESHRE, O-130). In this study, we evaluated (GCs) which affects reproductive outcome. Many shown a close association status GCs embryo’s quality. To date, with different protocols not been investigated. design, size, duration From November 2021 January 2022, all samples were obtained from provided written informed consent before inclusion cycles at single-institution. Single cell RNA-seq (scRNA-seq) performed on 2,224 corresponding MII-oocyte follicles total 16 normal reserve (aged < 40 years, AMH ≧1.1 ng/mL) undergoing chlormadinone acetate (n = 8), or cetrorelix 8). Participants/materials, setting, methods by follicular aspiration oocyte retrieval. fluid (FF) each follicle collected FF mixed together sequencing. isolated blood cellular debris using Percoll gradient centrifugation. Samples sequencing prepared Chromium Next GEM Cell V(D)J Reagent Kits. Libraries sequenced DNBSEQ-G400, reads processed CellRanger visualized CellxGene. Main results role chance Two population used investigation: MII-oocyte. After quality control, 1,197 1,027 GnRH-ant, respectively, analyzed. clustered into 6 groups based Leiden algorithm UMAP analysis. There no significant distribution cluster. two GCs, identified 12 differentially expressed (DEGs). Interestingly, observed increase mtDNA as data confirmed qRT-PCR Limitations, reasons caution Our conclusions are limited due Asian need be validated across centers other ethnicities. Due small sample these should prompt further confirm findings. Wider implications findings These describe, first time, DEGs investigated COS single-cell level. This suggests progestin elevation mt may associated outcomes. Trial registration number Not applicable