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Osteogenic cells form mineralized particles, a few μm in size, in a 3D collagen gel culture

Osteogenic cells form mineralized matrices in vitro, as well as in vivo. The formation and shape of the mineralized matrices are highly regulated by the cells. In vitro formation of mineralized matrices by osteogenic cells can be a model for in vivo osteogenesis. In this study, using a three-dimensi...

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Autores principales: Kihara, Takanori, Umezu, Chiya, Sawada, Karin, Furutani, Yukari
Formato: Online Artículo Texto
Lenguaje:English
Publicado: PeerJ Inc. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6815190/
https://www.ncbi.nlm.nih.gov/pubmed/31660270
http://dx.doi.org/10.7717/peerj.7889
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author Kihara, Takanori
Umezu, Chiya
Sawada, Karin
Furutani, Yukari
author_facet Kihara, Takanori
Umezu, Chiya
Sawada, Karin
Furutani, Yukari
author_sort Kihara, Takanori
collection PubMed
description Osteogenic cells form mineralized matrices in vitro, as well as in vivo. The formation and shape of the mineralized matrices are highly regulated by the cells. In vitro formation of mineralized matrices by osteogenic cells can be a model for in vivo osteogenesis. In this study, using a three-dimensional (3D) collagen gel culture system, we developed a new in vitro model for the formation of mineralized particles, a few µm in size, by the osteogenic cells. Human osteosarcoma (HOS) cells formed spherical mineralized matrices (about 12 µm) at approximately 7 days when cultured with β-glycerophosphate (β-GP)-containing culture media on 2D tissue culture plates. Alternately, when they were cultured in a 3D collagen gel containing β-GP, they formed mineralized particles with about 1.7 µm in the gel at approximately 3 days. Calcium precipitation in the gel was evaluated by measuring the gel turbidity. This type of mineralization of HOS cells, which formed mineralized particles inside the gel, was also observed in a peptide-based hydrogel culture. The mineralized particles were completely diminished by inhibiting the activity of Pit-1, phosphate cotransporter, of the HOS cells. When mouse osteoblast-like MC3T3-E1 cells, which form large and flat mineralized matrices in 2D osteogenic conditions at approximately 3 weeks of culture, were cultured in a 3D collagen gel, they also formed mineralized particles in the gel, similar to those in HOS cells, at approximately 18 days. Thus, osteogenic cells cultured in the 3D collagen gel form mineralized particles over a shorter period, and the mineralization could be easily determined by gel turbidity. This 3D gel culture system of osteogenic cells acts as a useful model for cells forming particle-type mineralized matrices, and we assume that the mineralized particles in the 3D hydrogel are calcospherulites, which are derived from matrix vesicles secreted by osteogenic cells.
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spelling pubmed-68151902019-10-28 Osteogenic cells form mineralized particles, a few μm in size, in a 3D collagen gel culture Kihara, Takanori Umezu, Chiya Sawada, Karin Furutani, Yukari PeerJ Bioengineering Osteogenic cells form mineralized matrices in vitro, as well as in vivo. The formation and shape of the mineralized matrices are highly regulated by the cells. In vitro formation of mineralized matrices by osteogenic cells can be a model for in vivo osteogenesis. In this study, using a three-dimensional (3D) collagen gel culture system, we developed a new in vitro model for the formation of mineralized particles, a few µm in size, by the osteogenic cells. Human osteosarcoma (HOS) cells formed spherical mineralized matrices (about 12 µm) at approximately 7 days when cultured with β-glycerophosphate (β-GP)-containing culture media on 2D tissue culture plates. Alternately, when they were cultured in a 3D collagen gel containing β-GP, they formed mineralized particles with about 1.7 µm in the gel at approximately 3 days. Calcium precipitation in the gel was evaluated by measuring the gel turbidity. This type of mineralization of HOS cells, which formed mineralized particles inside the gel, was also observed in a peptide-based hydrogel culture. The mineralized particles were completely diminished by inhibiting the activity of Pit-1, phosphate cotransporter, of the HOS cells. When mouse osteoblast-like MC3T3-E1 cells, which form large and flat mineralized matrices in 2D osteogenic conditions at approximately 3 weeks of culture, were cultured in a 3D collagen gel, they also formed mineralized particles in the gel, similar to those in HOS cells, at approximately 18 days. Thus, osteogenic cells cultured in the 3D collagen gel form mineralized particles over a shorter period, and the mineralization could be easily determined by gel turbidity. This 3D gel culture system of osteogenic cells acts as a useful model for cells forming particle-type mineralized matrices, and we assume that the mineralized particles in the 3D hydrogel are calcospherulites, which are derived from matrix vesicles secreted by osteogenic cells. PeerJ Inc. 2019-10-23 /pmc/articles/PMC6815190/ /pubmed/31660270 http://dx.doi.org/10.7717/peerj.7889 Text en ©2019 Kihara et al. https://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ) and either DOI or URL of the article must be cited.
spellingShingle Bioengineering
Kihara, Takanori
Umezu, Chiya
Sawada, Karin
Furutani, Yukari
Osteogenic cells form mineralized particles, a few μm in size, in a 3D collagen gel culture
title Osteogenic cells form mineralized particles, a few μm in size, in a 3D collagen gel culture
title_full Osteogenic cells form mineralized particles, a few μm in size, in a 3D collagen gel culture
title_fullStr Osteogenic cells form mineralized particles, a few μm in size, in a 3D collagen gel culture
title_full_unstemmed Osteogenic cells form mineralized particles, a few μm in size, in a 3D collagen gel culture
title_short Osteogenic cells form mineralized particles, a few μm in size, in a 3D collagen gel culture
title_sort osteogenic cells form mineralized particles, a few μm in size, in a 3d collagen gel culture
topic Bioengineering
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6815190/
https://www.ncbi.nlm.nih.gov/pubmed/31660270
http://dx.doi.org/10.7717/peerj.7889
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