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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PeerJ Inc.
2019-10-01
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| Online Access: | https://peerj.com/articles/7889.pdf |
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| author | Takanori Kihara Chiya Umezu Karin Sawada Yukari Furutani |
| author_facet | Takanori Kihara Chiya Umezu Karin Sawada Yukari Furutani |
| author_sort | Takanori Kihara |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-09T06:52:14Z |
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| language | English |
| last_indexed | 2024-03-09T06:52:14Z |
| publishDate | 2019-10-01 |
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| spelling | doaj.art-975787bf029746be9d28c064f1ae201f2023-12-03T10:24:57ZengPeerJ Inc.PeerJ2167-83592019-10-017e788910.7717/peerj.7889Osteogenic cells form mineralized particles, a few μm in size, in a 3D collagen gel cultureTakanori Kihara0Chiya Umezu1Karin Sawada2Yukari Furutani3Department of Life and Environment Engineering, Faculty of Environmental Engineering, The University of Kitakyushu, Kitakyushu, Fukuoka, JapanDepartment of Life and Environment Engineering, Faculty of Environmental Engineering, The University of Kitakyushu, Kitakyushu, Fukuoka, JapanDepartment of Life and Environment Engineering, Faculty of Environmental Engineering, The University of Kitakyushu, Kitakyushu, Fukuoka, JapanDepartment of Life and Environment Engineering, Faculty of Environmental Engineering, The University of Kitakyushu, Kitakyushu, Fukuoka, JapanOsteogenic 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.https://peerj.com/articles/7889.pdfMineralized matricesOsteogenic cellsCollagen gelMineralized particlesCalcospherulites3D culture |
| spellingShingle | Takanori Kihara Chiya Umezu Karin Sawada Yukari Furutani Osteogenic cells form mineralized particles, a few μm in size, in a 3D collagen gel culture PeerJ Mineralized matrices Osteogenic cells Collagen gel Mineralized particles Calcospherulites 3D 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 | Mineralized matrices Osteogenic cells Collagen gel Mineralized particles Calcospherulites 3D culture |
| url | https://peerj.com/articles/7889.pdf |
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