Using Patient-Derived Granulosa Cell Organoids as A Model System for Studying Granulosa Cell Dysfunction in Patients with Polycystic Ovary Syndrome (PCOS)

Abstract

Objective: To determine the feasibility of creating organoids from granulosa cells obtained at the time of oocyte retrieval and use these as a model system to investigate granulosa cell dysfunction in patients with polycystic ovary syndrome (PCOS).  

Design: Laboratory based study or prospective experimental study 

Materials and methods: Follicular fluid was collected from 44 patients at the time of oocyte retrieval. Granulosa cell organoids were created and used to examine granulosa cell dysfunction in patients with PCOS.  

Main Outcome Measures: The main outcome measure was the ability to grow granulosa cell organoids from granulosa cells obtained at the time of oocyte retrieval. Secondary outcome measures included growth of organoids, morphology, and defining the differentially expressed genes (DEGs) by bulk-RNA sequencing in patients with and without PCOS.  

Results: Granulosa cell organoids can be produced from granulosa cells obtained from the follicular fluid at the time of oocyte retrieval with a growth rate of 100% (44 patients). The presence of granulosa cells was confirmed by hematoxylin and eosin stains and confirmed by pathology. The morphology of granulosa cell organoids was examined by light microscopy and revealed a spherical pattern demonstrating 3D shape. Bulk RNA-sequencing was used to determine differences in gene expression in granulosa cells of patients with and without PCOS. Patients with PCOS had upregulated DEGs in TGF-B signaling, steroid metabolism, apoptosis and NOTCH signaling, and downregulation in luteinization, calcium channel and FGFR signaling.   

Conclusion: We report a novel protocol for development of granulosa cell organoids created from follicular fluid obtained at the time of oocyte retrieval. Significant differences in gene expression are shown from granulosa cell organoids between patients with and without PCOS in relation to apoptosis, folliculogenesis, and steroid metabolism. Granulosa cell organoids represent a novel experimental model system to gain more insight into the pathophysiology of PCOS.