# The emergence of multiple testicular cell lineages in human stem cell-derived testis-like organoids

## TL;DR

Scientists created testis-like organoids from human stem cells to study reproductive development and disorders.

## Contribution

The study provides the first comprehensive transcriptional profile of hiPSC-derived testis-like organoids.

## Key findings

- Transcriptomic analysis identified six distinct cell clusters resembling early testicular development.
- An inducible NR5A1/SF1 hiPSC line enhanced the emergence of Leydig and Sertoli cell markers.
- The organoids serve as a model system for studying differences of sex development in vitro.

## Abstract

Reproductive development is a complex process orchestrated by precise gene expression and cellular interactions. Disruption to this process can result in differences of sex development (DSDs) which occur in approximately 1-2% of live births. We have previously developed a protocol to differentiate human induced pluripotent stem cells (hiPSCs) into testis-like organoids. In this study, we performed bulk and single-cell RNA-sequencing on these organoids to investigate their transcriptional landscape. Transcriptomic analysis revealed six distinct cell clusters expressing markers associated with bipotential, early Sertoli and testicular interstitial cells. These findings provide the first comprehensive transcriptional profile of hiPSC-derived testis-like organoids. Additionally, to address the limited emergence of mature cell types, we generated an inducible NR5A1/SF1 hiPSC line, which successfully triggered the upregulation of Leydig cell markers and additional Sertoli markers upon overexpression. Our findings show that our testis-like organoids are a valuable model system for studying DSDs in vitro.

Summary: Human induced pluripotent stem cell-derived testis-like organoids contain distinct cell populations resembling early gonad development, offering a valuable model for studying human reproductive disorders.