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Gamblin, A. L., Renaud, A., Charrier, C., Hulin, P., Louarn, G., Heymann, D., Trichet, V. & Layrolle, P. (2014) Osteoblastic and osteoclastic differentiation of human mesenchymal stem cells and monocytes in a miniaturized three-dimensional culture with mineral granules. Acta Biomater. 10 5139–5147.
Added by: Laurent Cournède (2016-03-10 21:01:54) |
| Type de référence: Article DOI: 10.1016/j.actbio.2014.08.033 Numéro d'identification (ISBN etc.): 1742-7061 Clé BibTeX: Gamblin2014 Voir tous les détails bibliographiques  |
Catégories: PMN Mots-clés: 3-D model, Bone, bone regeneration, calcium-phosphate ceramics, coculture, disease, Human mesenchymal stromal/stem cells, Human monocytes, matrices, model, mouse, Osteoblasts, osteogenesis imperfecta, receptor, therapy Créateurs: Charrier, Gamblin, Heymann, Hulin, Layrolle, Louarn, Renaud, Trichet Collection: Acta Biomater. |
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| Résumé |
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The pathologies of the skeleton have a significant socioeconomic impact on our population. Although therapies have improved the treatment of osteosarcoma and osteoporosis, their efficacy still remains limited. In this context, we developed a miniaturized 3-D culture model of bone cells on calcium phosphate ceramics. Human bone marrow mesenchymal stem cells (MSCs) were three-dimensionally cultured on particles of biphasic calcium phosphate (BCP, 125-200 mu m) in osteogenic media. The MSCs seeded on the BCP particles adhered and proliferated, producing abundant collagenous extracellular matrix (ECM). Light and confocal laser scanning microscopy showed that the MSCs created bridges between the BCP particles and formed a 3-D structure. Energy dispersive X-ray analysis in a scanning electron microscope confirmed the mineralization of the collagen matrix. The 96-well sized bone constructs were tested by immunohistology and transcription analysis, proving cell differentiation. Both techniques corroborated the osteoblastic differentiation with high production of bone sialoprotein and osteocalcin. Peripheral blood CD14-positive monocytes (MOs) were pre-differentiated into osteoclasts prior to seeding on the 3-D constructs. Multinucleated and tartrate-resistant acid phosphatase-positive cells were also identified at the surface of the 3-D constructs after 90 days of culture. In addition, cell viability within these constructs was measured by flow cytometry. In summary, we have developed a miniaturized 3-D culture of bone cell precursors with osteoblasts and osteoclasts. This 3-D culture may make it possible to test the effects of new drugs for bone healing, osteoporosis and osteosarcomas, in more appropriate cell-cell and cell-matrix interactions than conventional 2-D cultures. (C) 2014 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.
Added by: Laurent Cournède |