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

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Harvard

Knarston, IM, Pachernegg, S, Robevska, G, Ghobrial, I, Er, PX, Georges, E, Takasato, M, Combes, AN, Jørgensen, A, Little, MH, Sinclair, AH & Ayers, KL 2020, 'An In Vitro Differentiation Protocol for Human Embryonic Bipotential Gonad and Testis Cell Development', Stem Cell Reviews and Reports, vol. 15, no. 6, pp. 1377-1391. https://doi.org/10.1016/j.stemcr.2020.10.009

APA

Knarston, I. M., Pachernegg, S., Robevska, G., Ghobrial, I., Er, P. X., Georges, E., Takasato, M., Combes, A. N., Jørgensen, A., Little, M. H., Sinclair, A. H., & Ayers, K. L. (2020). An In Vitro Differentiation Protocol for Human Embryonic Bipotential Gonad and Testis Cell Development. Stem Cell Reviews and Reports, 15(6), 1377-1391. https://doi.org/10.1016/j.stemcr.2020.10.009

CBE

Knarston IM, Pachernegg S, Robevska G, Ghobrial I, Er PX, Georges E, Takasato M, Combes AN, Jørgensen A, Little MH, Sinclair AH, Ayers KL. 2020. An In Vitro Differentiation Protocol for Human Embryonic Bipotential Gonad and Testis Cell Development. Stem Cell Reviews and Reports. 15(6):1377-1391. https://doi.org/10.1016/j.stemcr.2020.10.009

MLA

Vancouver

Author

Knarston, Ingrid M ; Pachernegg, Svenja ; Robevska, Gorjana ; Ghobrial, Irene ; Er, Pei Xuan ; Georges, Elizabeth ; Takasato, Minoru ; Combes, Alexander N ; Jørgensen, Anne ; Little, Melissa H ; Sinclair, Andrew H ; Ayers, Katie L. / An In Vitro Differentiation Protocol for Human Embryonic Bipotential Gonad and Testis Cell Development. In: Stem Cell Reviews and Reports. 2020 ; Vol. 15, No. 6. pp. 1377-1391.

Bibtex

@article{ef124228a10648aa9344ac52f41968b0,
title = "An In Vitro Differentiation Protocol for Human Embryonic Bipotential Gonad and Testis Cell Development",
abstract = "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.",
author = "Knarston, {Ingrid M} and Svenja Pachernegg and Gorjana Robevska and Irene Ghobrial and Er, {Pei Xuan} and Elizabeth Georges and Minoru Takasato and Combes, {Alexander N} and Anne J{\o}rgensen and Little, {Melissa H} and Sinclair, {Andrew H} and Ayers, {Katie L}",
note = "Copyright {\textcopyright} 2020 The Authors. Published by Elsevier Inc. All rights reserved.",
year = "2020",
month = dec,
day = "8",
doi = "10.1016/j.stemcr.2020.10.009",
language = "English",
volume = "15",
pages = "1377--1391",
journal = "Stem Cell Reviews and Reports",
issn = "1550-8943",
publisher = "Humana Press, Inc",
number = "6",

}

RIS

TY - JOUR

T1 - An In Vitro Differentiation Protocol for Human Embryonic Bipotential Gonad and Testis Cell Development

AU - Knarston, Ingrid M

AU - Pachernegg, Svenja

AU - Robevska, Gorjana

AU - Ghobrial, Irene

AU - Er, Pei Xuan

AU - Georges, Elizabeth

AU - Takasato, Minoru

AU - Combes, Alexander N

AU - Jørgensen, Anne

AU - Little, Melissa H

AU - Sinclair, Andrew H

AU - Ayers, Katie L

N1 - Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.

PY - 2020/12/8

Y1 - 2020/12/8

N2 - 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.

AB - 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.

U2 - 10.1016/j.stemcr.2020.10.009

DO - 10.1016/j.stemcr.2020.10.009

M3 - Journal article

C2 - 33217324

VL - 15

SP - 1377

EP - 1391

JO - Stem Cell Reviews and Reports

JF - Stem Cell Reviews and Reports

SN - 1550-8943

IS - 6

ER -

ID: 61412862