Characterization of Immortalized Osteoclast Precursors Developed from Mice Transgenic for Both bcl-XL and Simian Virus 40 Large T Antigen1
We recently developed an immortalized osteoclast (OCL) precursor cell line that forms large numbers of OCLs. This cell line was derived from mice doubly transgenic for bcl-XL and large T antigen that was targeted to cells in the OCL lineage (bcl-XL/Tag cells). We have now characterized these cells i...
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Published in | Endocrinology (Philadelphia) Vol. 140; no. 7; pp. 2954 - 2961 |
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Main Authors | , , , , , , |
Format | Journal Article |
Language | English |
Published |
Endocrine Society
01.07.1999
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Online Access | Get full text |
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Summary: | We recently developed an immortalized osteoclast (OCL) precursor cell
line that forms large numbers of OCLs. This cell line was derived from
mice doubly transgenic for bcl-XL and large
T antigen that was targeted to cells in the OCL lineage
(bcl-XL/Tag cells). We have now
characterized these cells in terms of their surface and enzymatic
phenotype, responsiveness to osteotropic factors, and differentiation
potential. The bcl-XL/Tag cells expressed
interleukin-1 receptors 1 and 2, gelatinase B (MMP9), as well as Mac-1,
CD16/CD32 (Fcγ receptors), CD45.2 (common leukocyte marker), CD86
(costimulatory molecule expressed on B cells, follicular dendritic
cells, and thymic epithelium), major histocompatibility complex I, and
nonspecific esterase when cocultured with MC3T3E1 cells. However, they
did not express the antigens for F4/80 (mature macrophage/dendritic
cell marker) by immunostaining. Treatment of
bcl-XL/Tag cells, cocultured with MC3T3E1
cells, with the combination of 1,25-dihydroxyvitamin D3 and
dexamethasone induced high levels of OCL formation. The
bcl-XL/Tag cells formed large numbers of
OCLs when cultured with RANK ligand and macrophage colony-stimulating
factor in the absence of feeder cells. In the absence of RANK ligand
and a feeder cell layer, 100% of the cells differentiated into
F4/80-positive cells. However, neither PTH nor PTH-related protein
enhanced OCL formation by bcl-XL/Tag cells
even when they were cocultured with primary osteoblasts, suggesting
that they differ from primary mouse bone marrow cells in their
responsiveness to PTH/PTH-related protein. Thus,
bcl-XL/Tag cells have many of the properties
of primary mouse OCL precursors and should be very useful for studies
of OCL differentiation and divergence of OCL precursors from the
macrophage lineage. |
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ISSN: | 0013-7227 1945-7170 |
DOI: | 10.1210/endo.140.7.6867 |