Cargando…

Fine‐tuning of dual‐SMAD inhibition to differentiate human pluripotent stem cells into neural crest stem cells

OBJECTIVES: The derivation of neural crest stem cells (NCSCs) from human pluripotent stem cells (hPSCs) has been commonly induced by WNT activation in combination with dual‐SMAD inhibition. In this study, by fine‐tuning BMP signalling in the conventional dual‐SMAD inhibition, we sought to generate l...

Descripción completa

Detalles Bibliográficos
Autores principales: Kim, Hyun‐Mun, Noh, Hye Bin, Lee, Sang‐Hyuk, Lee, Kun‐Gu, Chang, Bomi, Cheong, Eunji, Lee, C. Justin, Hwang, Dong‐Youn
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8450125/
https://www.ncbi.nlm.nih.gov/pubmed/34323338
http://dx.doi.org/10.1111/cpr.13103
_version_ 1784569561449758720
author Kim, Hyun‐Mun
Noh, Hye Bin
Lee, Sang‐Hyuk
Lee, Kun‐Gu
Chang, Bomi
Cheong, Eunji
Lee, C. Justin
Hwang, Dong‐Youn
author_facet Kim, Hyun‐Mun
Noh, Hye Bin
Lee, Sang‐Hyuk
Lee, Kun‐Gu
Chang, Bomi
Cheong, Eunji
Lee, C. Justin
Hwang, Dong‐Youn
author_sort Kim, Hyun‐Mun
collection PubMed
description OBJECTIVES: The derivation of neural crest stem cells (NCSCs) from human pluripotent stem cells (hPSCs) has been commonly induced by WNT activation in combination with dual‐SMAD inhibition. In this study, by fine‐tuning BMP signalling in the conventional dual‐SMAD inhibition, we sought to generate large numbers of NCSCs without WNT activation. MATERIALS AND METHODS: In the absence of WNT activation, we modulated the level of BMP signalling in the dual‐SMAD inhibition system to identify conditions that efficiently drove the differentiation of hPSCs into NCSCs. We isolated two NCSC populations separately and characterized them in terms of global gene expression profiles and differentiation ability. RESULTS: Our modified dual‐SMAD inhibition containing a lower dose of BMP inhibitor than that of the conventional dual‐SMAD inhibition drove hPSCs into mainly NCSCs, which consisted of HNK(+)p75high and HNK(+)p75low cell populations. We showed that the p75high population formed spherical cell clumps, while the p75low cell population generated a 2D monolayer. We detected substantial differences in gene expression profiles between the two cell groups and showed that both p75high and p75low cells differentiated into mesenchymal stem cells (MSCs), while only p75high cells had the ability to become peripheral neurons. CONCLUSIONS: This study will provide a framework for the generation and isolation of NCSC populations for effective cell therapy for peripheral neuropathies and MSC‐based cell therapy.
format Online
Article
Text
id pubmed-8450125
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher John Wiley and Sons Inc.
record_format MEDLINE/PubMed
spelling pubmed-84501252021-09-27 Fine‐tuning of dual‐SMAD inhibition to differentiate human pluripotent stem cells into neural crest stem cells Kim, Hyun‐Mun Noh, Hye Bin Lee, Sang‐Hyuk Lee, Kun‐Gu Chang, Bomi Cheong, Eunji Lee, C. Justin Hwang, Dong‐Youn Cell Prolif Original Articles OBJECTIVES: The derivation of neural crest stem cells (NCSCs) from human pluripotent stem cells (hPSCs) has been commonly induced by WNT activation in combination with dual‐SMAD inhibition. In this study, by fine‐tuning BMP signalling in the conventional dual‐SMAD inhibition, we sought to generate large numbers of NCSCs without WNT activation. MATERIALS AND METHODS: In the absence of WNT activation, we modulated the level of BMP signalling in the dual‐SMAD inhibition system to identify conditions that efficiently drove the differentiation of hPSCs into NCSCs. We isolated two NCSC populations separately and characterized them in terms of global gene expression profiles and differentiation ability. RESULTS: Our modified dual‐SMAD inhibition containing a lower dose of BMP inhibitor than that of the conventional dual‐SMAD inhibition drove hPSCs into mainly NCSCs, which consisted of HNK(+)p75high and HNK(+)p75low cell populations. We showed that the p75high population formed spherical cell clumps, while the p75low cell population generated a 2D monolayer. We detected substantial differences in gene expression profiles between the two cell groups and showed that both p75high and p75low cells differentiated into mesenchymal stem cells (MSCs), while only p75high cells had the ability to become peripheral neurons. CONCLUSIONS: This study will provide a framework for the generation and isolation of NCSC populations for effective cell therapy for peripheral neuropathies and MSC‐based cell therapy. John Wiley and Sons Inc. 2021-07-29 /pmc/articles/PMC8450125/ /pubmed/34323338 http://dx.doi.org/10.1111/cpr.13103 Text en © 2021 The Authors. Cell Proliferation published by John Wiley & Sons Ltd. https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Articles
Kim, Hyun‐Mun
Noh, Hye Bin
Lee, Sang‐Hyuk
Lee, Kun‐Gu
Chang, Bomi
Cheong, Eunji
Lee, C. Justin
Hwang, Dong‐Youn
Fine‐tuning of dual‐SMAD inhibition to differentiate human pluripotent stem cells into neural crest stem cells
title Fine‐tuning of dual‐SMAD inhibition to differentiate human pluripotent stem cells into neural crest stem cells
title_full Fine‐tuning of dual‐SMAD inhibition to differentiate human pluripotent stem cells into neural crest stem cells
title_fullStr Fine‐tuning of dual‐SMAD inhibition to differentiate human pluripotent stem cells into neural crest stem cells
title_full_unstemmed Fine‐tuning of dual‐SMAD inhibition to differentiate human pluripotent stem cells into neural crest stem cells
title_short Fine‐tuning of dual‐SMAD inhibition to differentiate human pluripotent stem cells into neural crest stem cells
title_sort fine‐tuning of dual‐smad inhibition to differentiate human pluripotent stem cells into neural crest stem cells
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8450125/
https://www.ncbi.nlm.nih.gov/pubmed/34323338
http://dx.doi.org/10.1111/cpr.13103
work_keys_str_mv AT kimhyunmun finetuningofdualsmadinhibitiontodifferentiatehumanpluripotentstemcellsintoneuralcreststemcells
AT nohhyebin finetuningofdualsmadinhibitiontodifferentiatehumanpluripotentstemcellsintoneuralcreststemcells
AT leesanghyuk finetuningofdualsmadinhibitiontodifferentiatehumanpluripotentstemcellsintoneuralcreststemcells
AT leekungu finetuningofdualsmadinhibitiontodifferentiatehumanpluripotentstemcellsintoneuralcreststemcells
AT changbomi finetuningofdualsmadinhibitiontodifferentiatehumanpluripotentstemcellsintoneuralcreststemcells
AT cheongeunji finetuningofdualsmadinhibitiontodifferentiatehumanpluripotentstemcellsintoneuralcreststemcells
AT leecjustin finetuningofdualsmadinhibitiontodifferentiatehumanpluripotentstemcellsintoneuralcreststemcells
AT hwangdongyoun finetuningofdualsmadinhibitiontodifferentiatehumanpluripotentstemcellsintoneuralcreststemcells