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  • SaOS2 Osteosarcoma cells as an in vitro model for studying the transition of human osteoblasts to osteocytes.

SaOS2 Osteosarcoma cells as an in vitro model for studying the transition of human osteoblasts to osteocytes.

Calcified tissue international (2014-06-12)
Matthew Prideaux, Asiri R Wijenayaka, Duminda D Kumarasinghe, Renee T Ormsby, Andreas Evdokiou, David M Findlay, Gerald J Atkins
ABSTRACT

The central importance of osteocytes in regulating bone homeostasis is becoming increasingly apparent. However, the study of these cells has been restricted by the relative paucity of cell line models, especially those of human origin. Therefore, we investigated the extent to which SaOS2 human osteosarcoma cells can differentiate into osteocyte-like cells. During culture under the appropriate mineralising conditions, SaOS2 cells reproducibly synthesised a bone-like mineralised matrix and temporally expressed the mature osteocyte marker genes SOST, DMP1, PHEX and MEPE and down-regulated expression of RUNX2 and COL1A1. SaOS2 cells cultured in 3D collagen gels acquired a dendritic morphology, characteristic of osteocytes, with multiple interconnecting cell processes. These findings suggest that SaOS2 cells have the capacity to differentiate into mature osteocyte-like cells under mineralising conditions. PTH treatment of SaOS2 cells resulted in strong down-regulation of SOST mRNA expression at all time points tested. Interestingly, PTH treatment resulted in the up-regulation of RANKL mRNA expression only at earlier stages of differentiation. These findings suggest that the response to PTH is dependent on the differentiation stage of the osteoblast/osteocyte. Together, our results demonstrate that SaOS2 cells can be used as a human model to investigate responses to osteotropic stimuli throughout differentiation to a mature osteocyte-like stage.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
NGF-β human, from human, recombinant, expressed in NSO cells, lyophilized powder, suitable for cell culture
Sigma-Aldrich
Cetylpyridinium chloride, meets USP testing specifications
USP
Cetylpyridinium chloride, United States Pharmacopeia (USP) Reference Standard
Cetylpyridinium chloride, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
DAPI, for nucleic acid staining