West China Journal of Stomatology

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Isolation of Bone Marrow-derived Rat Osteoclast-like Cells with the Digestion of Trypsin

DONG Qiang1, LIANG Xing2, CHEN Yue2, XU Ling1, ZHANG Qing-hong1, XIA Lu1, CHEN Ming1, FU Jun1   

  1. 1. Key. Laboratory of Oral Biomedical Engineering Ministry of Education, Sichuan University, Chengdu 610041, China; 2. Dept. of Prosthodontics, West China College of Stomatology, Sichuan University, Chengdu 610041, China
  • Received:2006-08-25 Revised:2006-08-25 Online:2006-08-20 Published:2006-08-20

Abstract:

 Objective  To obtain highly enriched osteoclasts in vitro. Methods  The 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. Results  The 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  teoclast-like cells were harvested by the digestion of 0.25% trypsin and 0.02% EDTA. Conclusion  It′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.

Key words: osteoclast, trypsin digestion, culture, purify