Isolation of bone marrow-derived rat osteoclast-like cells with the digestion of trypsin.
- Author:
Qiang DONG
1
;
Xing LIANG
;
Yue CHEN
;
Ling XU
;
Qing-hong ZHANG
;
Lu XIA
;
Ming CHEN
;
Jun FU
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Bone Marrow; Bone Marrow Cells; Bone and Bones; Calcitriol; Cells, Cultured; Dexamethasone; Osteoclasts; Rats; Rats, Sprague-Dawley; Trypsin
- From: West China Journal of Stomatology 2006;24(4):350-361
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo obtain highly enriched osteoclasts in vitro.
METHODSThe bone marrow cells of 1-day, 12-day, 24-day and 36-day old Sprague-Dawley (SD) rats were separately cultured with the presence of 1,25-(OH)2D3 and dexamethasone, and the osteoclast-like cells were identified by Trap staining and scanning electron microscope observation and were purified by trypsin digestion. The count of Trap positive osteoclast-like cells was analyzed statistically.
RESULTSThe osteoclast-like cells of each group were Trap positive cells and could form the bone absorption lacunas in vitro. The amounts of osteoclast-like cells were different statistically between the groups (P < 0.05) except between the groups of 1-day old and 12-day old (P > 0.05). Highly enriched osteoclast-like cells were harvested by the digestion of 0.25% trypsin and 0.02% EDTA.
CONCLUSIONIt's indicated that a large amount of highly enriched osteoclast-like cells could be obtained through the culture of bone marrow cells of 10-day and 12-day old SD rats with 1,25-(OH)2D3 and dexamethasone and the digestion of trypsin/EDTA.
