Characterization of immortalized osteoclast precursors developed from mice transgenic for both bcl-X(L) and simian virus 40 large T antigen

Teuvo A. Hentunen, Sharon H. Jackson, Hoyeon Chung, Sakamuri V. Reddy, Joseph Lorenzo, Sun Jin Choi, G. David Roodman

Research output: Contribution to journalArticle

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Abstract

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-X(L) and large T antigen that was targeted to cells in the OCL lineage (bcl-X(L)/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-X(L)/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 histocompatibiiity 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-X(L)/Tag cells, cocultured with MC3T3E1 cells, with the combination of 1,25-dihydroxyvitamin D3 and dexamethasone induced high levels of OCL formation. The bcl-X(L)/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-X(L)/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-X(L)/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.

Original languageEnglish (US)
Pages (from-to)2954-2961
Number of pages8
JournalEndocrinology
Volume140
Issue number7
StatePublished - 1999
Externally publishedYes

Fingerprint

Simian virus 40
Viral Tumor Antigens
Osteoclasts
Transgenic Mice
RANK Ligand
Feeder Cells
Macrophages
Follicular Dendritic Cells
Cell Line
Carboxylesterase
Macrophage Colony-Stimulating Factor
Interleukin-1 Receptors
Fc Receptors
Calcitriol
Matrix Metalloproteinase 9
Osteoblasts
Bone Marrow Cells
Dendritic Cells
Dexamethasone
Proteins

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Hentunen, T. A., Jackson, S. H., Chung, H., Reddy, S. V., Lorenzo, J., Choi, S. J., & Roodman, G. D. (1999). Characterization of immortalized osteoclast precursors developed from mice transgenic for both bcl-X(L) and simian virus 40 large T antigen. Endocrinology, 140(7), 2954-2961.

Characterization of immortalized osteoclast precursors developed from mice transgenic for both bcl-X(L) and simian virus 40 large T antigen. / Hentunen, Teuvo A.; Jackson, Sharon H.; Chung, Hoyeon; Reddy, Sakamuri V.; Lorenzo, Joseph; Choi, Sun Jin; Roodman, G. David.

In: Endocrinology, Vol. 140, No. 7, 1999, p. 2954-2961.

Research output: Contribution to journalArticle

Hentunen, TA, Jackson, SH, Chung, H, Reddy, SV, Lorenzo, J, Choi, SJ & Roodman, GD 1999, 'Characterization of immortalized osteoclast precursors developed from mice transgenic for both bcl-X(L) and simian virus 40 large T antigen', Endocrinology, vol. 140, no. 7, pp. 2954-2961.
Hentunen, Teuvo A. ; Jackson, Sharon H. ; Chung, Hoyeon ; Reddy, Sakamuri V. ; Lorenzo, Joseph ; Choi, Sun Jin ; Roodman, G. David. / Characterization of immortalized osteoclast precursors developed from mice transgenic for both bcl-X(L) and simian virus 40 large T antigen. In: Endocrinology. 1999 ; Vol. 140, No. 7. pp. 2954-2961.
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abstract = "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-X(L) and large T antigen that was targeted to cells in the OCL lineage (bcl-X(L)/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-X(L)/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 histocompatibiiity 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-X(L)/Tag cells, cocultured with MC3T3E1 cells, with the combination of 1,25-dihydroxyvitamin D3 and dexamethasone induced high levels of OCL formation. The bcl-X(L)/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-X(L)/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-X(L)/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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AU - Reddy, Sakamuri V.

AU - Lorenzo, Joseph

AU - Choi, Sun Jin

AU - Roodman, G. David

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