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Cell passage and composition of culture medium effects proliferation and differentiation of human osteoblast-like cells from facial bone.
Pradel, W; Mai, R; Gedrange, T; Lauer, G.
Afiliação
  • Pradel W; Department of Oral and Maxillofacial Surgery, University Hospital Carl Gustav Carus Dresden Technical University Dresden, Germany. Germany.
J Physiol Pharmacol ; 59 Suppl 5: 47-58, 2008 Nov.
Article em En | MEDLINE | ID: mdl-19075324
ABSTRACT
Cells loose their capability to multiply and to differentiate when they are serial subcultivated. However, both, multiplication and differentiation are of utmost importance to obtain sufficient amounts of cells for the translation of tissue regeneration into cell based therapeutic approaches. Thus, for the clinical application more information about ideal culture conditions are necessary. Therefore, aim of this study was to assess culture conditions of human osteoblast-like cells during long-term culture focusing on effects of different culture media and ascorbic acid. Biopsies of maxilla and mandible were obtained from 17 patients to test different cell culture media and from 10 patients to analyse differentiation and proliferation related to number of subcultures and ascorbic acid content. Histochemical and immunhistochemical tests (EZ4U assay, ALP histochemistry, type I collagen immunohistochemistry, osteocalcin Elisa) were performed to determine cell proliferation and differentiation. Opti-MEM with 10% FCS produced statistically significant the highest increase in cell counts. The highest proliferation rate in long-term cultivation was seen in the 4th cell passage. A reciprocal relationship between cell proliferation and differentiation over 5 passages with a turning point in the 4(th) passage was found. An ascorbic acid content of 50 microg/ml triggered an optimal increase in differentiation. For osteoblast-like cells, Opti-MEM with 10% FCS proved to be the best culture medium. After 3 passages there is the highest amount of cells with osteogenic differentiation which is enhanced by the addition of ascorbic acid. This approach is suitable for tissue engineering of bone grafts.
Assuntos
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Osteoblastos / Ácido Ascórbico / Diferenciação Celular / Meios de Cultura / Proliferação de Células / Ossos Faciais Limite: Adolescent / Adult / Aged / Aged80 / Child / Female / Humans / Male / Middle aged Idioma: En Revista: J Physiol Pharmacol Assunto da revista: FARMACOLOGIA / FISIOLOGIA Ano de publicação: 2008 Tipo de documento: Article
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Osteoblastos / Ácido Ascórbico / Diferenciação Celular / Meios de Cultura / Proliferação de Células / Ossos Faciais Limite: Adolescent / Adult / Aged / Aged80 / Child / Female / Humans / Male / Middle aged Idioma: En Revista: J Physiol Pharmacol Assunto da revista: FARMACOLOGIA / FISIOLOGIA Ano de publicação: 2008 Tipo de documento: Article