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Initial cell plating density affects properties of human primary synovial mesenchymal stem cells
Synovial mesenchymal stem cells (MSCs) appear to be an attractive cell source in cartilage and meniscus regeneration because of their high proliferative and chondrogenic potentials. Two methods are used to culture synovial nucleated cells in the preparation of primary synovial MSCs. In one method, t...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6585790/ https://www.ncbi.nlm.nih.gov/pubmed/30035340 http://dx.doi.org/10.1002/jor.24112 |
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author | Nakamura, Kaori Tsuji, Kunikazu Mizuno, Mitsuru Koga, Hideyuki Muneta, Takeshi Sekiya, Ichiro |
author_facet | Nakamura, Kaori Tsuji, Kunikazu Mizuno, Mitsuru Koga, Hideyuki Muneta, Takeshi Sekiya, Ichiro |
author_sort | Nakamura, Kaori |
collection | PubMed |
description | Synovial mesenchymal stem cells (MSCs) appear to be an attractive cell source in cartilage and meniscus regeneration because of their high proliferative and chondrogenic potentials. Two methods are used to culture synovial nucleated cells in the preparation of primary synovial MSCs. In one method, the cells are plated at low density to make cell colonies. In the other method, the cells are plated at high density. We investigated the effects of initial cell density on proliferation, surface markers, and multipotentiality, including chondrogenesis in primary synovial MSCs. Human synovium was obtained from the knee joints of patients with osteoarthritis after total knee arthroplasty. Immediately after enzyme digestion, the synovial nucleated cells were plated in densities of 10(3), 10(4), or 10(5) cells/60‐cm(2) dish and cultured for 14 days. Proliferation, surface markers, chondrogenesis, adipogenesis, and calcification were examined in three populations. The cell colonies were distinct in the 10(3) cells/dish group, faint in the 10(4) cells/dish group, and obscure in the 10(5) cells/dish group. The total number of cells/dish was positively related to plating density, whereas the fold increase was negatively related to plating density (n = 13). Among 12 surface markers, a negative relation to plating density was distinct in CD105. The cartilage pellet weight was negatively related to the initial plating density. The oil red‐o positive area and alizarin red positive area were positively related to the initial plating density. The initial cell plating density affected the properties of primary synovial MSCs. Synovial nucleated cells proliferated better when plated at low density, and the synovial MSCs obtained by this method contained a high chondrogenic potential. © 2018 The Authors. Journal of Orthopaedic Research® Published by Wiley Periodicals, Inc. J Orthop Res 37:1358–1367, 2019. |
format | Online Article Text |
id | pubmed-6585790 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-65857902019-06-27 Initial cell plating density affects properties of human primary synovial mesenchymal stem cells Nakamura, Kaori Tsuji, Kunikazu Mizuno, Mitsuru Koga, Hideyuki Muneta, Takeshi Sekiya, Ichiro J Orthop Res Research Articles Synovial mesenchymal stem cells (MSCs) appear to be an attractive cell source in cartilage and meniscus regeneration because of their high proliferative and chondrogenic potentials. Two methods are used to culture synovial nucleated cells in the preparation of primary synovial MSCs. In one method, the cells are plated at low density to make cell colonies. In the other method, the cells are plated at high density. We investigated the effects of initial cell density on proliferation, surface markers, and multipotentiality, including chondrogenesis in primary synovial MSCs. Human synovium was obtained from the knee joints of patients with osteoarthritis after total knee arthroplasty. Immediately after enzyme digestion, the synovial nucleated cells were plated in densities of 10(3), 10(4), or 10(5) cells/60‐cm(2) dish and cultured for 14 days. Proliferation, surface markers, chondrogenesis, adipogenesis, and calcification were examined in three populations. The cell colonies were distinct in the 10(3) cells/dish group, faint in the 10(4) cells/dish group, and obscure in the 10(5) cells/dish group. The total number of cells/dish was positively related to plating density, whereas the fold increase was negatively related to plating density (n = 13). Among 12 surface markers, a negative relation to plating density was distinct in CD105. The cartilage pellet weight was negatively related to the initial plating density. The oil red‐o positive area and alizarin red positive area were positively related to the initial plating density. The initial cell plating density affected the properties of primary synovial MSCs. Synovial nucleated cells proliferated better when plated at low density, and the synovial MSCs obtained by this method contained a high chondrogenic potential. © 2018 The Authors. Journal of Orthopaedic Research® Published by Wiley Periodicals, Inc. J Orthop Res 37:1358–1367, 2019. John Wiley and Sons Inc. 2018-08-16 2019-06 /pmc/articles/PMC6585790/ /pubmed/30035340 http://dx.doi.org/10.1002/jor.24112 Text en © 2018 The Authors. Journal of Orthopaedic Research® Published by Wiley Periodicals, Inc. This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made. |
spellingShingle | Research Articles Nakamura, Kaori Tsuji, Kunikazu Mizuno, Mitsuru Koga, Hideyuki Muneta, Takeshi Sekiya, Ichiro Initial cell plating density affects properties of human primary synovial mesenchymal stem cells |
title | Initial cell plating density affects properties of human primary synovial mesenchymal stem cells |
title_full | Initial cell plating density affects properties of human primary synovial mesenchymal stem cells |
title_fullStr | Initial cell plating density affects properties of human primary synovial mesenchymal stem cells |
title_full_unstemmed | Initial cell plating density affects properties of human primary synovial mesenchymal stem cells |
title_short | Initial cell plating density affects properties of human primary synovial mesenchymal stem cells |
title_sort | initial cell plating density affects properties of human primary synovial mesenchymal stem cells |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6585790/ https://www.ncbi.nlm.nih.gov/pubmed/30035340 http://dx.doi.org/10.1002/jor.24112 |
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