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| ISOLATION AND CHARACTERIZATION OF HIGHLY ENRICHED, PREFUSION MOUSE OSTEOCLASTIC CELLS |
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| Author(s): WESOLOWSKI G, DUONG LT, LAKKAKORPI PT, NAGY RM, TEZUKA K, TANAKA H, RODAN GA, RODAN SB |
| Source: EXPERIMENTAL CELL RESEARCH Volume: 219 Issue: 2 Pages: 679-686 Published: AUG 1995 |
| Times Cited: 71 References: 51 |
| Abstract: 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 cm(2) 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 I-125-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 pp60(c-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)(2)D-3. 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. (C) 1995 Academic Press, Inc. |
| Document Type: Article |
| Language: English |
Addresses:
1. MERCK SHARP & DOHME LTD, DEPT BONE BIOL & OSTEOPOROSIS RES, RES LABS, W POINT, PA 19486 USA |
| Publisher: ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS, 525B STREET, SUITE 1900, SAN DIEGO, CA 92101-4495 |
| Subject Category: Oncology; Cell Biology |
| IDS Number: RQ063 |
| ISSN: 0014-4827 |
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