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Simple and Robust Differentiation of Human Pluripotent Stem Cells toward Chondrocytes by Two Small-Molecule Compounds
A simple induction protocol to differentiate chondrocytes from pluripotent stem cells (PSCs) using small-molecule compounds is beneficial for cartilage regenerative medicine and mechanistic studies of chondrogenesis. Here, we demonstrate that chondrocytes are robustly induced from human PSCs by simp...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6739881/ https://www.ncbi.nlm.nih.gov/pubmed/31402337 http://dx.doi.org/10.1016/j.stemcr.2019.07.012 |
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author | Kawata, Manabu Mori, Daisuke Kanke, Kosuke Hojo, Hironori Ohba, Shinsuke Chung, Ung-il Yano, Fumiko Masaki, Hideki Otsu, Makoto Nakauchi, Hiromitsu Tanaka, Sakae Saito, Taku |
author_facet | Kawata, Manabu Mori, Daisuke Kanke, Kosuke Hojo, Hironori Ohba, Shinsuke Chung, Ung-il Yano, Fumiko Masaki, Hideki Otsu, Makoto Nakauchi, Hiromitsu Tanaka, Sakae Saito, Taku |
author_sort | Kawata, Manabu |
collection | PubMed |
description | A simple induction protocol to differentiate chondrocytes from pluripotent stem cells (PSCs) using small-molecule compounds is beneficial for cartilage regenerative medicine and mechanistic studies of chondrogenesis. Here, we demonstrate that chondrocytes are robustly induced from human PSCs by simple combination of two compounds, CHIR99021, a glycogen synthase kinase 3 inhibitor, and TTNPB, a retinoic acid receptor (RAR) agonist, under serum- and feeder-free conditions within 5–9 days. An excellent differentiation efficiency and potential to form hyaline cartilaginous tissues in vivo were demonstrated. Comprehensive gene expression and open chromatin analyses at each protocol stage revealed step-by-step differentiation toward chondrocytes. Genome-wide analysis of RAR and β-catenin association with DNA showed that retinoic acid and Wnt/β-catenin signaling collaboratively regulated the key marker genes at each differentiation stage. This method provides a promising cell source for regenerative medicine and, as an in vitro model, may facilitate elucidation of the molecular mechanisms underlying chondrocyte differentiation. |
format | Online Article Text |
id | pubmed-6739881 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-67398812019-09-16 Simple and Robust Differentiation of Human Pluripotent Stem Cells toward Chondrocytes by Two Small-Molecule Compounds Kawata, Manabu Mori, Daisuke Kanke, Kosuke Hojo, Hironori Ohba, Shinsuke Chung, Ung-il Yano, Fumiko Masaki, Hideki Otsu, Makoto Nakauchi, Hiromitsu Tanaka, Sakae Saito, Taku Stem Cell Reports Article A simple induction protocol to differentiate chondrocytes from pluripotent stem cells (PSCs) using small-molecule compounds is beneficial for cartilage regenerative medicine and mechanistic studies of chondrogenesis. Here, we demonstrate that chondrocytes are robustly induced from human PSCs by simple combination of two compounds, CHIR99021, a glycogen synthase kinase 3 inhibitor, and TTNPB, a retinoic acid receptor (RAR) agonist, under serum- and feeder-free conditions within 5–9 days. An excellent differentiation efficiency and potential to form hyaline cartilaginous tissues in vivo were demonstrated. Comprehensive gene expression and open chromatin analyses at each protocol stage revealed step-by-step differentiation toward chondrocytes. Genome-wide analysis of RAR and β-catenin association with DNA showed that retinoic acid and Wnt/β-catenin signaling collaboratively regulated the key marker genes at each differentiation stage. This method provides a promising cell source for regenerative medicine and, as an in vitro model, may facilitate elucidation of the molecular mechanisms underlying chondrocyte differentiation. Elsevier 2019-08-08 /pmc/articles/PMC6739881/ /pubmed/31402337 http://dx.doi.org/10.1016/j.stemcr.2019.07.012 Text en © 2019 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Article Kawata, Manabu Mori, Daisuke Kanke, Kosuke Hojo, Hironori Ohba, Shinsuke Chung, Ung-il Yano, Fumiko Masaki, Hideki Otsu, Makoto Nakauchi, Hiromitsu Tanaka, Sakae Saito, Taku Simple and Robust Differentiation of Human Pluripotent Stem Cells toward Chondrocytes by Two Small-Molecule Compounds |
title | Simple and Robust Differentiation of Human Pluripotent Stem Cells toward Chondrocytes by Two Small-Molecule Compounds |
title_full | Simple and Robust Differentiation of Human Pluripotent Stem Cells toward Chondrocytes by Two Small-Molecule Compounds |
title_fullStr | Simple and Robust Differentiation of Human Pluripotent Stem Cells toward Chondrocytes by Two Small-Molecule Compounds |
title_full_unstemmed | Simple and Robust Differentiation of Human Pluripotent Stem Cells toward Chondrocytes by Two Small-Molecule Compounds |
title_short | Simple and Robust Differentiation of Human Pluripotent Stem Cells toward Chondrocytes by Two Small-Molecule Compounds |
title_sort | simple and robust differentiation of human pluripotent stem cells toward chondrocytes by two small-molecule compounds |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6739881/ https://www.ncbi.nlm.nih.gov/pubmed/31402337 http://dx.doi.org/10.1016/j.stemcr.2019.07.012 |
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