Document Detail

Isolation and characterization of highly enriched, prefusion mouse osteoclastic cells.
MedLine Citation:
PMID:  7641819     Owner:  NLM     Status:  MEDLINE    
This study describes the isolation and characterization of highly enriched mammalian osteoclast precursors, released by the "disintegrin" echistatin, from an osteoclast formation culture. Incubation of a 6-day coculture of mouse bone marrow cells and mouse osteoblastic cells (MB1.8) with echistatin (30 nM), an RGD-containing snake venom, for 20 min yielded an 88-95% pure population of tartrate-resistant acid phosphatase-positive cells, 1.5 x 10(5) cells per 150 cm2 culture dish. These cells were mostly mononucleated and based on the following characteristics are considered to be prefusion osteoclasts (pOC cells): (i) presence of calcitonin (CT) receptors documented by 125I-sCT autoradiography and cAMP generation in response to salmon calcitonin; (ii) expression of mRNAs for alpha v beta 3 integrin, osteopontin, 92-kDa type IV collagenase (matrix metalloproteinase 9), carbonic anhydrase II, OC-2 (an "osteoclastic" cysteine proteinase), and protein tyrosine phosphatase epsilon; and (iii) high level expression of pp60c-src protein. The pOC cells resorb bone (form "pits" on bone slices) but only in the presence of osteoblastic MB1.8 cells and 1,25(OH)2D3. Resorption was inhibited by CT. In conclusion, we describe a rapid, reproducible procedure to isolate virtually pure mammalian prefusion osteoclasts, which should help in the study of osteoclast formation, composition, and function.
G Wesolowski; L T Duong; P T Lakkakorpi; R M Nagy; K Tezuka; H Tanaka; G A Rodan; S B Rodan
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Experimental cell research     Volume:  219     ISSN:  0014-4827     ISO Abbreviation:  Exp. Cell Res.     Publication Date:  1995 Aug 
Date Detail:
Created Date:  1995-09-18     Completed Date:  1995-09-18     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0373226     Medline TA:  Exp Cell Res     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  679-86     Citation Subset:  IM    
Department of Bone Biology and Osteoporosis Research, Merck Research Laboratories, West Point, Pennsylvania 19486, USA.
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MeSH Terms
Bone Marrow Cells*
Cell Separation / methods*
Cells, Cultured
Integrins / biosynthesis
Osteoclasts / cytology*,  metabolism
Peptides / pharmacology
Sialoglycoproteins / biosynthesis
Viper Venoms / pharmacology
Reg. No./Substance:
0/Integrins; 0/Peptides; 0/Sialoglycoproteins; 0/Spp1 protein, mouse; 0/Viper Venoms; 106441-73-0/Osteopontin; 118337-11-4/echistatin

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