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Udgivet

An In Vitro Differentiation Protocol for Human Embryonic Bipotential Gonad and Testis Cell Development

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  • Ingrid M Knarston
  • Svenja Pachernegg
  • Gorjana Robevska
  • Irene Ghobrial
  • Pei Xuan Er
  • Elizabeth Georges
  • Minoru Takasato
  • Alexander N Combes
  • Anne Jørgensen
  • Melissa H Little
  • Andrew H Sinclair
  • Katie L Ayers
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Currently an in vitro model that fully recapitulates the human embryonic gonad is lacking. Here we describe a fully defined feeder-free protocol to generate early testis-like cells with the ability to be cultured as an organoid, from human induced pluripotent stem cells. This stepwise approach uses small molecules to mimic embryonic development, with upregulation of bipotential gonad markers (LHX9, EMX2, GATA4, and WT1) at day 10 of culture, followed by induction of testis Sertoli cell markers (SOX9, WT1, and AMH) by day 15. Aggregation into 3D structures and extended culture on Transwell filters yielded organoids with defined tissue structures and distinct Sertoli cell marker expression. These studies provide insight into human gonadal development, suggesting that a population of precursor cells may originate from a more lateral region of the mesoderm. Our protocol represents a significant advance toward generating a much-needed human gonad organoid for studying disorders/differences of sex development.

OriginalsprogEngelsk
TidsskriftStem Cell Reviews and Reports
Vol/bind15
Udgave nummer6
Sider (fra-til)1377-1391
Antal sider15
ISSN1550-8943
DOI
StatusUdgivet - 8 dec. 2020

Bibliografisk note

Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.

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