RRC ID |
41485
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著者 |
Quinn JM, Whitty GA, Byrne RJ, Gillespie MT, Hamilton JA.
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タイトル |
The generation of highly enriched osteoclast-lineage cell populations.
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ジャーナル |
Bone
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Abstract |
Osteoclasts form when hematopoietic cells are stimulated by macrophage colony-stimulating factor (M-CSF) and receptor activator of NF-kappaB ligand (RANKL) or tumor necrosis factor-alpha (TNFalpha). Osteoclast precursors derive from M-CSF-dependent proliferating hematopoietic cells but cannot yet be purified from mixed populations. M-CSF stimulation of bone marrow cells results in large numbers of nonadherent, proliferating macrophage precursors. These rapidly form adherent bone marrow macrophages (BMM). BMM and their precursors can be isolated free from mesenchymal and lymphocytic cells. BMM precursors derived from CBA-strain mouse bone marrow, when cocultured with ST2 cells (which express RANKL and M-CSF), formed numerous mononuclear osteoclasts, which resorbed bone and expressed tartrate-resistant acid phosphatase (TRAP) and calcitonin receptors (CTR). Addition of approximately 10 BMM precursors to ST2 cultures resulted in over 80% of these cocultures forming functional osteoclasts, suggesting that they are a highly enriched source of osteoclast progenitors. Supporting this, recombinant RANKL/M-CSF-stimulated BMM precursors formed populations in which all cells expressed TRAP. While only a small proportion of these cells (8.6%) expressed CTR, with transforming growth factor-beta (TGFbeta) present RANKL/M-CSF-stimulated BMM precursors formed almost pure (98.4%) CTR-positive osteoclasts after 7 days. This suggests that TGFbeta stimulated the maturation rate of these cells. Passaged or viably frozen BMM precursors gave rise to BMM that also all formed osteoclasts lineage cells after RANKL/M-CSF stimulation. These data suggest that BMM precursors derived from CBA mice are an expanded pool of osteoclast progenitors. These can be employed to generate osteoclast populations of high purity and in large numbers when stimulated by TGFbeta, which greatly augments the osteoclastogenic effects of RANKL.
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巻・号 |
30(1)
|
ページ |
164-70
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公開日 |
2002-1-1
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DOI |
10.1016/s8756-3282(01)00654-8
|
PII |
S8756328201006548
|
PMID |
11792580
|
MeSH |
Animals
Bone Marrow Cells / cytology*
Bone Marrow Cells / drug effects
Bone Resorption / etiology
Bone Resorption / pathology
Carrier Proteins / pharmacology
Cell Differentiation / drug effects
Cell Line
Coculture Techniques
Hematopoietic Stem Cells / cytology
Hematopoietic Stem Cells / drug effects
Macrophage Colony-Stimulating Factor / pharmacology
Membrane Glycoproteins / pharmacology
Mice
Mice, Inbred C57BL
Mice, Inbred CBA
Osteoclasts / cytology*
Osteoclasts / drug effects
RANK Ligand
Receptor Activator of Nuclear Factor-kappa B
Recombinant Proteins / pharmacology
Tumor Necrosis Factor-alpha / pharmacology
|
IF |
4.147
|
引用数 |
50
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WOS 分野
|
ENDOCRINOLOGY & METABOLISM
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リソース情報 |
ヒト・動物細胞 |
|