West China Journal of Stomatology 2006;24(4):350-361

Isolation of bone marrow-derived rat osteoclast-like cells with the digestion of trypsin.

Qiang DONG 1 ; Xing LIANG ; Yue CHEN ; Ling XU ; Qing-hong ZHANG ; Lu XIA ; Ming CHEN ; Jun FU

Affiliations

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Country

China

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.