Osteogenic differentiation of immature osteoblasts: Interplay of cell culture media and supplements

Biotechnic & Histochemistry : Official Publication of the Biological Stain Commission
A BrauerW Metzger

Abstract

Differentiation of immature osteoblasts to mature osteoblasts in vitro initially was induced by supplementing the medium with β-gylcerophosphate and dexamethasone. Later, ascorbic acid, vitamin D3, vitamin K3 and TGFβ1 were used in varying concentrations as supplements to generate a mature osteoblast phenotype. We tested the effects of several combinations of cell culture media, seeding protocols and osteogenic supplements on osteogenic differentiation of human primary osteoblasts. Osteogenic differentiation was analyzed by staining alkaline phosphatase (ALP) with 5-bromo-4-chloro-3-indolyl-phosphate/nitro blue tetrazolium (BCIP/NBT) and by von Kossa staining of deposited calcium phosphate. The combinations of culture media and supplements significantly influenced osteogenic differentiation, but the seeding protocol did not. Staining of ALP and calcium phosphate could be achieved only if our own mix of osteogenic supplements was used in combination with Dulbecco's modified Eagle medium or if a commercial mix of osteogenic supplements was used in combination with osteoblast growth medium. Especially for von Kossa, we observed great variations in the staining intensity. Because osteogenic differentiation is a complex process, the...Continue Reading

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Citations

Jul 26, 2016·Frontiers in Cellular and Infection Microbiology·Jérôme JosseSophie C Gangloff
Oct 11, 2016·Oral Surgery, Oral Medicine, Oral Pathology and Oral Radiology·Philipp KauffmannRudolf Matthias Gruber
Nov 20, 2016·Journal of Cellular and Molecular Medicine·Guo-Ping ZhangGuo-Bin Liu
May 4, 2018·Journal of Materials Science. Materials in Medicine·O HasturkV Hasirci
Oct 2, 2020·Journal of Orthopaedic Research : Official Publication of the Orthopaedic Research Society·Michael B BergerBarbara D Boyan
Jul 7, 2020·Journal of Agricultural and Food Chemistry·Divya UpadhyaySuman Kapila

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