Document Detail


Isolation and characterization of highly enriched, prefusion mouse osteoclastic cells.
MedLine Citation:
PMID:  7641819     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
G Wesolowski; L T Duong; P T Lakkakorpi; R M Nagy; K Tezuka; H Tanaka; G A Rodan; S B Rodan
Related Documents :
18664329 - Internal ribosome entry site of rhopalosiphum padi virus is functional in mammalian cel...
5890139 - Preservation of mammalian cells in a chemically defined medium and dimethylsulfoxide.
20014139 - Enhancing glycoprotein sialylation by targeted gene silencing in mammalian cells.
7300029 - A comparative study of assay systems for drug-resistant mutation in cultured mammalian ...
19640839 - Role of mammalian ecdysoneless in cell cycle regulation.
1368109 - A novel micromanipulation technique for measuring the bursting strength of single mamma...
7523029 - The photodynamic occlusion of choroidal vessels using benzoporphyrin derivative.
24691869 - Time-lapse video microscopy and image analysis of adherence and growth patterns of cand...
2836939 - Mechanisms of cell killing/cytopathic effects by nonhuman retroviruses.
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    
Affiliation:
Department of Bone Biology and Osteoporosis Research, Merck Research Laboratories, West Point, Pennsylvania 19486, USA.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Animals
Bone Marrow Cells*
Cell Separation / methods*
Cells, Cultured
Integrins / biosynthesis
Mice
Osteoclasts / cytology*,  metabolism
Osteopontin
Peptides / pharmacology
Sialoglycoproteins / biosynthesis
Viper Venoms / pharmacology
Chemical
Reg. No./Substance:
0/Integrins; 0/Peptides; 0/Sialoglycoproteins; 0/Spp1 protein, mouse; 0/Viper Venoms; 106441-73-0/Osteopontin; 118337-11-4/echistatin

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Widely used enhancer of eukaryotic expression vectors is strongly and differentially regulated in fi...
Next Document:  Lamin dynamics during sea urchin male pronuclear formation in vitro.