A Novel Osteogenic Cell Line That Differentiates Into GFP ‐Tagged Osteocytes and Forms Mineral With a Bone‐Like Lacunocanalicular Structure

ABSTRACTOsteocytes, the most abundant cells in bone, were once thought to be inactive, but are now known to have multifunctional roles in bone, including in mechanotransduction, regulation of osteoblast and osteoclast function and phosphate homeostasis. Because osteocytes are embedded in a mineralized matrix and are challenging to study, there is a need for new tools and cell models to understand their biology. We have generated two clonal osteogenic cell lines, OmGFP66 and OmGFP10, by immortalization of primary bone cells from mice expressing a membrane ‐targeted GFP driven by theDmp1‐promoter. One of these clones, OmGFP66, has unique properties compared with previous osteogenic and osteocyte cell models and forms 3‐dimensional mineralized bone‐like structures, containing highly dendritic GFP‐positive osteocytes, embedded in clearly defined lacunae. Confocal and electron microscopy showed that structurally and morphologically, these bone‐like structures resemble bone in vivo, even mimicking the lacunocanalicular ultrastructure and 3D spacing of in vivo osteocytes. In osteogenic conditions, OmGFP66 cells express alkaline phosphatase (ALP), produce a mineralized ty pe I collagen matrix, and constitutively express the early osteocyte marker,E11/gp38. With differentiation they express osteocyte markers,Dmp1, Phex, Mepe, Fgf23, and the mature osteocyte marker,Sost. They also expressRankL, Opg, andHif1 α, and show expected osteocyte responses to PTH, including downreg...
Source: Journal of Bone and Mineral Research - Category: Orthopaedics Authors: Tags: ORIGINAL ARTICLE Source Type: research