Cell Lines

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Brief Summary: UMKC Researchers have developed novel cell lines that are useful in the examination of osteocyte function, biomineralization, SOST/sclerostin, FGF23 and other mechanisms of osteoblast-to-osteocyte differentiation. Detailed Description: The two cell lines were isolated from long bone of a mouse that was generated by crossing the Immortomouse® with a mouse where the DMP1 promoter drives expression of the GFP. One of the cell lines, IDG-SW3 (SW3), expresses all the markers of osteocytes including Dmp1-GFP, Dmp1, E11/gp38, SOST/sclerostin, and FGF23.  The second cell line, IDG-TI (T1), mainly expresses the characteristics of the matrix producing osteoblast such as high alkaline phosphatase, with delayed expression of Dmp1-GFP and E11/gp38, but no expression of SOST/sclerostin or FGF23.  Both cells will produce new bone in vivo.  Uses:  To generate large numbers of osteocyte-like cells to produce sufficient quantities of osteocytes for study. To generate large numbers of cells of a homogeneous stage of osteogenic differentiation. To study osteocyte secretion of sclerostin, such as screening for sclerostin antagonists. To investigate regulation of FGF23 expression in osteocytes and the role of osteocytes in regulation blood calcium/phosphate homeostasis. To study the role of osteocytes as mechano-sensory cells and their role in regulating bone response to mechanical stress. To screen potential new therapies to induce bone formation. To track cells responsible for bone formation in vivo. To identify additional osteocyte-selective markers and receptors.  Advantages: This invention is an improvement over previous cell lines due to the following factors: 1. The cells are maintained in a non-differentiated state at 33°C in the presence of interferon- g (IFN-g), which allows large scale production without the loss of phenotype as occurs with other cell lines; 2. Upon culture at 37°C in the absence of IFN-g, the temperature-sensitive large T-antigen is no longer expressed, no longer functional, and no longer contributes to the cell phenotype. Thus, the cells have the same gene expression as primary cells; 3. The cells are clonal, so all cells are homogeneous and at the same stage of differentiation; 4. The IDG-SW3 cells express the series of markers of the early-to-late osteocyte including Dmp1-GFP, E11/gp38, SOST/sclerostin, and FGF23; 5. These cells can be maintained not only in 2D cultures but also in 3D cultures; 6. These cells are viable up to 35-50 days; and 7. These cells will generate new bone in vivo.