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Modulation of osteogenic differentiation in human skeletal cells in Vitro by 5-azacytidine

Modulation of osteogenic differentiation in human skeletal cells in Vitro by 5-azacytidine
Modulation of osteogenic differentiation in human skeletal cells in Vitro by 5-azacytidine
Cellular differentiation is controlled by a variety of factors including gene methylation, which represses particular genes as cell fate is determined. The incorporation of 5-azacytidine (5azaC) into DNA in vitro prevents methylation and thus can alter cellular differentiation pathways. Human bone marrow fibroblasts and MG63 cells treated with 5azaC were used as models of osteogenic progenitors and of a more mature osteoblast phenotype, respectively. The capacity for differentiation of these cells following treatment with glucocorticoids was investigated. 5azaC treatment led to significant expression of the osteoblastic marker alkaline phosphatase in MG63 osteosarcoma cells, which was further augmented by glucocorticoids; however, in human marrow fibroblasts alkaline phosphatase activity was only observed in glucocorticoid-treated cultures. MG63 cells represent a phenotype late in the osteogenic lineage in which demethylation is sufficient to induce alkaline phosphatase activity. Marrow fibroblasts are at an earlier stage of differentiation and require stimulation with glucocorticoids. In contrast, the expression of osteocalcin, an osteoblastic marker, was unaffected by 5azaC treatment, suggesting that regulation of expression of the osteocalcin gene does not involve methylation. These models provide novel approaches to the study of the control of differentiation in the marrow fibroblastic system.
1065-6995
207-15
Locklin, R.M.
879f95db-0fc5-4353-a81b-9aec5d24bc5e
Oreffo, R.O.
ff9fff72-6855-4d0f-bfb2-311d0e8f3778
Triffitt, J.T.
06d3019a-06e6-4abd-9e73-f073d621e1f9
Locklin, R.M.
879f95db-0fc5-4353-a81b-9aec5d24bc5e
Oreffo, R.O.
ff9fff72-6855-4d0f-bfb2-311d0e8f3778
Triffitt, J.T.
06d3019a-06e6-4abd-9e73-f073d621e1f9

Locklin, R.M., Oreffo, R.O. and Triffitt, J.T. (1998) Modulation of osteogenic differentiation in human skeletal cells in Vitro by 5-azacytidine. Cell Biology International, 22 (3), 207-15. (doi:10.1006/cbir.1998.0240). (PMID:9974215)

Record type: Article

Abstract

Cellular differentiation is controlled by a variety of factors including gene methylation, which represses particular genes as cell fate is determined. The incorporation of 5-azacytidine (5azaC) into DNA in vitro prevents methylation and thus can alter cellular differentiation pathways. Human bone marrow fibroblasts and MG63 cells treated with 5azaC were used as models of osteogenic progenitors and of a more mature osteoblast phenotype, respectively. The capacity for differentiation of these cells following treatment with glucocorticoids was investigated. 5azaC treatment led to significant expression of the osteoblastic marker alkaline phosphatase in MG63 osteosarcoma cells, which was further augmented by glucocorticoids; however, in human marrow fibroblasts alkaline phosphatase activity was only observed in glucocorticoid-treated cultures. MG63 cells represent a phenotype late in the osteogenic lineage in which demethylation is sufficient to induce alkaline phosphatase activity. Marrow fibroblasts are at an earlier stage of differentiation and require stimulation with glucocorticoids. In contrast, the expression of osteocalcin, an osteoblastic marker, was unaffected by 5azaC treatment, suggesting that regulation of expression of the osteocalcin gene does not involve methylation. These models provide novel approaches to the study of the control of differentiation in the marrow fibroblastic system.

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Published date: 1998
Organisations: Human Development & Health

Identifiers

Local EPrints ID: 360861
URI: https://eprints.soton.ac.uk/id/eprint/360861
ISSN: 1065-6995
PURE UUID: 601f84e6-fa07-4e7c-8d08-ae39cc9efb18
ORCID for R.O. Oreffo: ORCID iD orcid.org/0000-0001-5995-6726

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Date deposited: 09 Jan 2014 08:46
Last modified: 06 Jun 2018 12:53

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