title: [Separation of osteoclasts by lectin affinity chromatography].
abstract: Newborn rat calvaria bone cells obtained by digestion were fractionated on columns of wheat-germ agglutinin (WGA) sepharose 6MB for osteoclast isolation. The initial nonspecific binding cells which were passed through the WGA sepharose column by a buffer acquired a high enzyme activity of alkaline phosphatase, but not that of acid phosphatase. However, elution of cells using a buffer with the addition of N-acetyl-D-glucosamine resulted in a high acid phosphatase activity but no alkaline phosphatase activity. The former WGA binding negative fraction enriched osteoblasts averaging 30 microns in size. The latter WGA binding positive fraction enriched osteoclasts ranging from 20 microns to 60 microns in size. The electron-microscope clearly demonstrated the cellular details of osteoclasts. Isolated cell counts showed a ratio of six to four. These results indicate that our method of osteoclast isolation is simple and useful in lectin affinity chromatography because all cells have sugar moieties on their surface and the binding of osteoclasts can be reversed by the addition of specific lectin-binding sugars to the eluting buffer.
authors.affiliation: Department of Orthopaedic Surgery, Kinki University School of Medicine, Osaka, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Itokazu
authors.forename: A
authors.initials: A
authors.lastname: Tan
authors.forename: S
authors.initials: S
authors.lastname: Tanaka
mesh_terms: D000135:Acid Phosphatase / Q000378:metabolism; D000818:Animals; D002469:Cell Separation / Q000379:methods*; D002846:Chromatography, Affinity; D037102:Lectins; D010010:Osteoclasts / Q000378:metabolism / Q000648:ultrastructure; D051381:Rats; D011919:Rats, Inbred Strains
chemical_list: D037102:Lectins; D000135:Acid Phosphatase