Directed Differentiation of Human IPSCs to Functional Ovarian Granulosa-like Cells Via Transcription Factor Overexpression

Overview

Journal Elife

Specialty Biology

Date 2023 Feb 21

PMID 36803359

Authors

Merrick D Pierson Smela

Christian C Kramme

Patrick R J Fortuna

Jessica L Adams

Rui Su

Edward Dong

Mutsumi Kobayashi

Garyk Brixi

Venkata Srikar Kavirayuni

Emma Tysinger

Richie E Kohman

Toshi Shioda

Pranam Chatterjee

George M Church

Affiliations

Soon will be listed here.

Abstract

An in vitro model of human ovarian follicles would greatly benefit the study of female reproduction. Ovarian development requires the combination of germ cells and several types of somatic cells. Among these, granulosa cells play a key role in follicle formation and support for oogenesis. Whereas efficient protocols exist for generating human primordial germ cell-like cells (hPGCLCs) from human induced pluripotent stem cells (hiPSCs), a method of generating granulosa cells has been elusive. Here, we report that simultaneous overexpression of two transcription factors (TFs) can direct the differentiation of hiPSCs to granulosa-like cells. We elucidate the regulatory effects of several granulosa-related TFs and establish that overexpression of NR5A1 and either RUNX1 or RUNX2 is sufficient to generate granulosa-like cells. Our granulosa-like cells have transcriptomes similar to human fetal ovarian cells and recapitulate key ovarian phenotypes including follicle formation and steroidogenesis. When aggregated with hPGCLCs, our cells form ovary-like organoids (ovaroids) and support hPGCLC development from the premigratory to the gonadal stage as measured by induction of DAZL expression. This model system will provide unique opportunities for studying human ovarian biology and may enable the development of therapies for female reproductive health.

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