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Dynamic formation of cellular aggregates of chondrocytes and mesenchymal stem cells in spinner flask

OBJECTIVES: Cellular aggregates are readily applicable in cell‐based therapy. The effects of agitation and inoculation density on the aggregation of cells in spinner flask and the molecular mechanism of aggregation were investigated. MATERIALS AND METHODS: The aggregation kinetics of cells in spinne...

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Autores principales: He, Huimin, He, Qing, Xu, Feiyue, Zhou, Yan, Ye, Zhaoyang, Tan, Wen‐Song
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6669002/
https://www.ncbi.nlm.nih.gov/pubmed/31206838
http://dx.doi.org/10.1111/cpr.12587
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author He, Huimin
He, Qing
Xu, Feiyue
Zhou, Yan
Ye, Zhaoyang
Tan, Wen‐Song
author_facet He, Huimin
He, Qing
Xu, Feiyue
Zhou, Yan
Ye, Zhaoyang
Tan, Wen‐Song
author_sort He, Huimin
collection PubMed
description OBJECTIVES: Cellular aggregates are readily applicable in cell‐based therapy. The effects of agitation and inoculation density on the aggregation of cells in spinner flask and the molecular mechanism of aggregation were investigated. MATERIALS AND METHODS: The aggregation kinetics of cells in spinner flask was evaluated with bovine articular chondrocytes (bACs), rabbit bone marrow‐derived mesenchymal stem cells (rMSCs) and their mixture. The morphology of cellular aggregates was studied with scanning electron microscopy and gene expression of cell adhesion‐related molecules was analysed. RESULTS: It was shown that suspension culture in spinner flask induced the aggregation of bACs and rMSCs. Both cells exhibited increased aggregation rate and aggregate size with decreasing agitation rate and increasing cell inoculation density. Additionally, aggregate size increased with extended culture time. By analysing gene expression of integrin β1 and cadherin, it was indicated that these molecules were potentially involved in the aggregation process of bACs and rMSCs, respectively. Aggregates composed of both bACs and rMSCs were also prepared, showing rMSCs in the core and bACs in the periphery. CONCLUSIONS: Cellular aggregates were prepared in dynamic suspension culture using spinner flask, the key parameters to the aggregation process were identified, and the molecular mechanism of aggregation was revealed. This would lay a solid foundation for the large‐scale production of cellular aggregates for cell‐based therapy, such as cartilage regeneration.
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spelling pubmed-66690022020-03-13 Dynamic formation of cellular aggregates of chondrocytes and mesenchymal stem cells in spinner flask He, Huimin He, Qing Xu, Feiyue Zhou, Yan Ye, Zhaoyang Tan, Wen‐Song Cell Prolif Original Articles OBJECTIVES: Cellular aggregates are readily applicable in cell‐based therapy. The effects of agitation and inoculation density on the aggregation of cells in spinner flask and the molecular mechanism of aggregation were investigated. MATERIALS AND METHODS: The aggregation kinetics of cells in spinner flask was evaluated with bovine articular chondrocytes (bACs), rabbit bone marrow‐derived mesenchymal stem cells (rMSCs) and their mixture. The morphology of cellular aggregates was studied with scanning electron microscopy and gene expression of cell adhesion‐related molecules was analysed. RESULTS: It was shown that suspension culture in spinner flask induced the aggregation of bACs and rMSCs. Both cells exhibited increased aggregation rate and aggregate size with decreasing agitation rate and increasing cell inoculation density. Additionally, aggregate size increased with extended culture time. By analysing gene expression of integrin β1 and cadherin, it was indicated that these molecules were potentially involved in the aggregation process of bACs and rMSCs, respectively. Aggregates composed of both bACs and rMSCs were also prepared, showing rMSCs in the core and bACs in the periphery. CONCLUSIONS: Cellular aggregates were prepared in dynamic suspension culture using spinner flask, the key parameters to the aggregation process were identified, and the molecular mechanism of aggregation was revealed. This would lay a solid foundation for the large‐scale production of cellular aggregates for cell‐based therapy, such as cartilage regeneration. John Wiley and Sons Inc. 2019-06-17 /pmc/articles/PMC6669002/ /pubmed/31206838 http://dx.doi.org/10.1111/cpr.12587 Text en © 2019 The Authors. Cell Proliferation Published by John Wiley & Sons Ltd. This is an open access article under the terms of the http://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
He, Huimin
He, Qing
Xu, Feiyue
Zhou, Yan
Ye, Zhaoyang
Tan, Wen‐Song
Dynamic formation of cellular aggregates of chondrocytes and mesenchymal stem cells in spinner flask
title Dynamic formation of cellular aggregates of chondrocytes and mesenchymal stem cells in spinner flask
title_full Dynamic formation of cellular aggregates of chondrocytes and mesenchymal stem cells in spinner flask
title_fullStr Dynamic formation of cellular aggregates of chondrocytes and mesenchymal stem cells in spinner flask
title_full_unstemmed Dynamic formation of cellular aggregates of chondrocytes and mesenchymal stem cells in spinner flask
title_short Dynamic formation of cellular aggregates of chondrocytes and mesenchymal stem cells in spinner flask
title_sort dynamic formation of cellular aggregates of chondrocytes and mesenchymal stem cells in spinner flask
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6669002/
https://www.ncbi.nlm.nih.gov/pubmed/31206838
http://dx.doi.org/10.1111/cpr.12587
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