| Isolation of avian osteoclasts: improved techniques to preferentially purify viable cells. | |
| | |
MedLine Citation:
|
PMID: 1858522 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
Among the many different methods that have been used to obtain and study isolated osteoclasts from a variety of species, the egg-laying hen maintained on a low-calcium diet has proven to be one of the richest sources of relatively large numbers of osteoclasts. However, recent reports and our own observations indicate that only a very small proportion of the osteoclasts harvested by such methods are viable. The difficulty in obtaining large numbers of viable osteoclasts has restricted studies of osteoclast function and regulation, and so new isolation methods were sought. This report describes an osteoclast isolation procedure designed to substantially enrich for large numbers of viable authentic osteoclasts. Size and cell density differences between osteoclasts and contaminating mononuclear cells have been exploited in developing the methods for osteoclast enrichment. Sequential nonenzymatic and enzymatic procedures, followed by cell density separations, have yielded three populations of osteoclasts derived from chick hatchlings maintained on a low-calcium diet. A corresponding decrease in bone-associated osteoclasts during the sequential isolation scheme has been monitored using an osteoclast-directed monoclonal antibody, 121F. The first two populations contain 40% osteoclasts, which are predominantly (greater than 99%) nonviable, but the third population contains 8-fold more viable osteoclasts, effectively increasing the proportion of viable osteoclasts more than 25-fold in comparison with the first two populations. The osteoclast-like nature of the isolated viable population 3 cells was established by demonstrating ruffled border formation, possession of the 121F monoclonal antibody-reactive osteoclast antigen, bone particle resorption activity, and resorption pit formation on cortical bone slices revealed by transmission and scanning electron microscopy. |
| | |
Authors:
|
M J Oursler; P Collin-Osdoby; F Anderson; L Li; D Webber; P Osdoby |
Publication Detail:
|
Type: Journal Article; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
|
Title: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research Volume: 6 ISSN: 0884-0431 ISO Abbreviation: J. Bone Miner. Res. Publication Date: 1991 Apr |
Date Detail:
|
Created Date: 1991-08-28 Completed Date: 1991-08-28 Revised Date: 2007-11-14 |
Medline Journal Info:
|
Nlm Unique ID: 8610640 Medline TA: J Bone Miner Res Country: UNITED STATES |
Other Details:
|
Languages: eng Pagination: 375-85 Citation Subset: IM |
Affiliation:
|
Department of Cell Biology, Washington University School of Dental Medicine, St. Louis, MO. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Animals Bone Resorption / pathology Cell Count Cell Separation / methods* Cell Survival / physiology Cells, Cultured Chickens Enzyme-Linked Immunosorbent Assay Microscopy, Electron Microscopy, Electron, Scanning Osteoclasts / cytology* |
| Grant Support | |
ID/Acronym/Agency:
|
AR 32087/AR/NIAMS NIH HHS; AR 32927/AR/NIAMS NIH HHS; K04-AM1474/AM/NIADDK NIH HHS |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Effect of chronic carbonic anhydrase inhibitor therapy on bone mineral density in white women.
Next Document: A systemic acceleratory phenomenon (SAP) accompanies the regional acceleratory phenomenon (RAP) duri...