Journal article

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

Ingrid M Knarston, Svenja Pachernegg, Gorjana Robevska, Irene Ghobrial, Pei Xuan Er, Elizabeth Georges, Minoru Takasato, Alexander N Combes, Anne Jorgensen, Melissa H Little, Andrew H Sinclair, Katie L Ayers



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. Th..

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Awarded by National Health and Medical Research Council (NHMRC), Australia

Funding Acknowledgements

This project was funded by an NHMRC Ideas grant (K.L.A., APP1156942) and Program grant (A.H.S., APP1074258) from the National Health and Medical Research Council (NHMRC), Australia. I.M.K was funded by a Research Training Scheme scholarship, The University of Melbourne, and an International Foundation for Ethical Research (IFER) graduate fellowship. M.H.L. and A.H.S. are Senior Principal Research Fellows of the NHMRC, Australia (APP1136085 and APP1154187 respectively).