Osteoblasts isolated from mouse calvaria initiate matrix mineralization in culture - PubMed (original) (raw)
Osteoblasts isolated from mouse calvaria initiate matrix mineralization in culture
B Ecarot-Charrier et al. J Cell Biol. 1983 Mar.
Abstract
A method is presented for isolating osteoblasts from newborn mouse calvaria without the use of digestive enzymes. The procedure is based on the ability of osteoblasts to migrate from bone onto small glass fragments (Jones, S.J., and A. Boyde, 1977, Cell Tissue Res., 184:179-193). The isolated cells were cultured for up to 14 d in Dulbecco's modified Eagle's medium supplemented with 10% fetal calf serum and 50 micrograms/ml of ascorbic acid. 7-d cultures were incubated for 24 h with [3H]proline. High levels of collagen synthesis relative to total protein were found, as measured by collagenase digestion of medium and cell layer proteins. Analysis of pepsin-digested proteins from the same cultures by SDS PAGE showed that type I collagen was predominantly produced with small amounts of type III and V (alpha 1 chains) collagens. Osteoblasts grown in the presence of beta-glycerophosphate were able to initiate mineral deposition in culture. Electron microscopic analysis of the cultures revealed the presence of needle-shaped apatite-like crystals associated with collagen fibrils and vesicles in the extracellular space. Mouse skin fibroblasts cultured under identical conditions failed to initiate mineralization. Electron histochemical studies revealed the presence of alkaline phosphatase activity, associated with osteoblast membranes, matrix vesicles and on or near collagen fibrils. Thus these isolated osteoblasts retained in culture their unique property of initiating mineralization and therefore represent a model of value for studying the mineralization process in vitro.
Similar articles
- Mineralization in vitro of matrix formed by osteoblasts isolated by collagenase digestion.
Nefussi JR, Boy-Lefevre ML, Boulekbache H, Forest N. Nefussi JR, et al. Differentiation. 1985;29(2):160-8. doi: 10.1111/j.1432-0436.1985.tb00310.x. Differentiation. 1985. PMID: 2995186 - Characterization of endosteal osteoblastic cells isolated from mouse caudal vertebrae.
Lomri A, Marie PJ, Tran PV, Hott M. Lomri A, et al. Bone. 1988;9(3):165-75. doi: 10.1016/8756-3282(88)90006-3. Bone. 1988. PMID: 2844214 - Distinct proliferative and differentiated stages of murine MC3T3-E1 cells in culture: an in vitro model of osteoblast development.
Quarles LD, Yohay DA, Lever LW, Caton R, Wenstrup RJ. Quarles LD, et al. J Bone Miner Res. 1992 Jun;7(6):683-92. doi: 10.1002/jbmr.5650070613. J Bone Miner Res. 1992. PMID: 1414487 - von Kossa staining alone is not sufficient to confirm that mineralization in vitro represents bone formation.
Bonewald LF, Harris SE, Rosser J, Dallas MR, Dallas SL, Camacho NP, Boyan B, Boskey A. Bonewald LF, et al. Calcif Tissue Int. 2003 May;72(5):537-47. doi: 10.1007/s00223-002-1057-y. Epub 2003 May 6. Calcif Tissue Int. 2003. PMID: 12724828 - In vitro osteogenesis assays: influence of the primary cell source on alkaline phosphatase activity and mineralization.
Hoemann CD, El-Gabalawy H, McKee MD. Hoemann CD, et al. Pathol Biol (Paris). 2009 Jun;57(4):318-23. doi: 10.1016/j.patbio.2008.06.004. Epub 2008 Oct 7. Pathol Biol (Paris). 2009. PMID: 18842361 Review.
Cited by
- Calcification of osteoblastlike rat osteosarcoma cells in agarose suspension cultures.
Nishimoto SK, Stryker WF, Nimni ME. Nishimoto SK, et al. Calcif Tissue Int. 1987 Nov;41(5):274-80. doi: 10.1007/BF02555229. Calcif Tissue Int. 1987. PMID: 3121151 - Formation of a new fibrous attachment to human dental roots. Effect of co-culturing periodontal ligament-derived and allogenic cortical bone-derived cells.
Bernstein AB, Preisig E, Schroeder HE. Bernstein AB, et al. Cell Tissue Res. 1989 Mar;255(3):631-9. doi: 10.1007/BF00218801. Cell Tissue Res. 1989. PMID: 2706664 - Monolayer cultures of normal human bone cells contain multiple subpopulations of alkaline phosphatase positive cells.
Matsuyama T, Lau KH, Wergedal JE. Matsuyama T, et al. Calcif Tissue Int. 1990 Nov;47(5):276-83. doi: 10.1007/BF02555909. Calcif Tissue Int. 1990. PMID: 2257520 - Human bone cells in vitro.
Robey PG, Termine JD. Robey PG, et al. Calcif Tissue Int. 1985 Sep;37(5):453-60. Calcif Tissue Int. 1985. PMID: 2998572 - Biomimetic systems for hydroxyapatite mineralization inspired by bone and enamel.
Palmer LC, Newcomb CJ, Kaltz SR, Spoerke ED, Stupp SI. Palmer LC, et al. Chem Rev. 2008 Nov;108(11):4754-83. doi: 10.1021/cr8004422. Chem Rev. 2008. PMID: 19006400 Free PMC article. Review. No abstract available.
References
- J Cell Biol. 1982 Feb;92(2):452-61 - PubMed
- Nature. 1970 Aug 15;227(5259):680-5 - PubMed
- Calcif Tissue Int. 1982 Jan;34(1):76-9 - PubMed
- Calcif Tissue Int. 1982 May;34(3):291-4 - PubMed
- Eur J Biochem. 1974 Jul 1;46(1):83-8 - PubMed
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources