title: SaOS2 Osteosarcoma cells as an in vitro model for studying the transition of human osteoblasts to osteocytes.
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.
authors.affiliation: Bone Cell Biology Group, Centre for Orthopaedic and Trauma Research, The University of Adelaide, North Terrace, Adelaide, SA, 5005, Australia.
authors.forename: Matthew
authors.initials: M
authors.lastname: Prideaux
authors.forename: Asiri R
authors.initials: AR
authors.lastname: Wijenayaka
authors.forename: Duminda D
authors.initials: DD
authors.lastname: Kumarasinghe
authors.forename: Renee T
authors.initials: RT
authors.lastname: Ormsby
authors.forename: Andreas
authors.initials: A
authors.lastname: Evdokiou
authors.forename: David M
authors.initials: DM
authors.lastname: Findlay
authors.forename: Gerald J
authors.initials: GJ
authors.lastname: Atkins
mesh_terms: D002113:Calcification, Physiologic / Q000502:physiology*; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology*; D045744:Cell Line, Tumor / Q000166:cytology; D006801:Humans; D066298:In Vitro Techniques; D018613:Microscopy, Confocal; D010006:Osteoblasts / Q000166:cytology* / Q000378:metabolism; D010011:Osteocytes / Q000166:cytology* / Q000378:metabolism; D012516:Osteosarcoma; D060888:Real-Time Polymerase Chain Reaction; D013052:Spectrometry, X-Ray Emission