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Transcriptome-Wide Analysis of Human Chondrocyte Expansion on Synoviocyte Matrix

Human chondrocytes are expanded and used in autologous chondrocyte implantation techniques and are known to rapidly de-differentiate in culture. These chondrocytes, when cultured on tissue culture plastic (TCP), undergo both phenotypical and morphological changes and quickly lose the ability to re-d...

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Autores principales: Kean, Thomas J., Ge, Zhongqi, Li, Yumei, Chen, Rui, Dennis, James E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6406362/
https://www.ncbi.nlm.nih.gov/pubmed/30678371
http://dx.doi.org/10.3390/cells8020085
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author Kean, Thomas J.
Ge, Zhongqi
Li, Yumei
Chen, Rui
Dennis, James E.
author_facet Kean, Thomas J.
Ge, Zhongqi
Li, Yumei
Chen, Rui
Dennis, James E.
author_sort Kean, Thomas J.
collection PubMed
description Human chondrocytes are expanded and used in autologous chondrocyte implantation techniques and are known to rapidly de-differentiate in culture. These chondrocytes, when cultured on tissue culture plastic (TCP), undergo both phenotypical and morphological changes and quickly lose the ability to re-differentiate to produce hyaline-like matrix. Growth on synoviocyte-derived extracellular matrix (SDECM) reduces this de-differentiation, allowing for more than twice the number of population doublings (PD) whilst retaining chondrogenic capacity. The goal of this study was to apply RNA sequencing (RNA-Seq) analysis to examine the differences between TCP-expanded and SDECM-expanded human chondrocytes. Human chondrocytes from three donors were thawed from primary stocks and cultured on TCP flasks or on SDECM-coated flasks at physiological oxygen tension (5%) for 4 passages. During log expansion, RNA was extracted from the cell layer (70–90% confluence) at passages 1 and 4. Total RNA was column-purified and DNAse-treated before quality control analysis and next-generation RNA sequencing. Significant effects on gene expression were observed due to both culture surface and passage number. These results offer insight into the mechanism of how SDECM provides a more chondrogenesis-preserving environment for cell expansion, the transcriptome-wide changes that occur with culture, and potential mechanisms for further enhancement of chondrogenesis-preserving growth.
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spelling pubmed-64063622019-03-19 Transcriptome-Wide Analysis of Human Chondrocyte Expansion on Synoviocyte Matrix Kean, Thomas J. Ge, Zhongqi Li, Yumei Chen, Rui Dennis, James E. Cells Article Human chondrocytes are expanded and used in autologous chondrocyte implantation techniques and are known to rapidly de-differentiate in culture. These chondrocytes, when cultured on tissue culture plastic (TCP), undergo both phenotypical and morphological changes and quickly lose the ability to re-differentiate to produce hyaline-like matrix. Growth on synoviocyte-derived extracellular matrix (SDECM) reduces this de-differentiation, allowing for more than twice the number of population doublings (PD) whilst retaining chondrogenic capacity. The goal of this study was to apply RNA sequencing (RNA-Seq) analysis to examine the differences between TCP-expanded and SDECM-expanded human chondrocytes. Human chondrocytes from three donors were thawed from primary stocks and cultured on TCP flasks or on SDECM-coated flasks at physiological oxygen tension (5%) for 4 passages. During log expansion, RNA was extracted from the cell layer (70–90% confluence) at passages 1 and 4. Total RNA was column-purified and DNAse-treated before quality control analysis and next-generation RNA sequencing. Significant effects on gene expression were observed due to both culture surface and passage number. These results offer insight into the mechanism of how SDECM provides a more chondrogenesis-preserving environment for cell expansion, the transcriptome-wide changes that occur with culture, and potential mechanisms for further enhancement of chondrogenesis-preserving growth. MDPI 2019-01-24 /pmc/articles/PMC6406362/ /pubmed/30678371 http://dx.doi.org/10.3390/cells8020085 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kean, Thomas J.
Ge, Zhongqi
Li, Yumei
Chen, Rui
Dennis, James E.
Transcriptome-Wide Analysis of Human Chondrocyte Expansion on Synoviocyte Matrix
title Transcriptome-Wide Analysis of Human Chondrocyte Expansion on Synoviocyte Matrix
title_full Transcriptome-Wide Analysis of Human Chondrocyte Expansion on Synoviocyte Matrix
title_fullStr Transcriptome-Wide Analysis of Human Chondrocyte Expansion on Synoviocyte Matrix
title_full_unstemmed Transcriptome-Wide Analysis of Human Chondrocyte Expansion on Synoviocyte Matrix
title_short Transcriptome-Wide Analysis of Human Chondrocyte Expansion on Synoviocyte Matrix
title_sort transcriptome-wide analysis of human chondrocyte expansion on synoviocyte matrix
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6406362/
https://www.ncbi.nlm.nih.gov/pubmed/30678371
http://dx.doi.org/10.3390/cells8020085
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