[Isolation of bone marrow-derived rat osteoclast-like cells with the digestion of trypsin].
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 osteoclast-like cells were harvested by the digestion of 0.25% trypsin and 0.02% EDTA.


CONCLUSIONS
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.
Dong
Qiang
Q
Key Laboratory of Oral Biomedical Engineering, Ministry of Education, Sichuan University, Chengdu 610041, China.
Liang
Xing
X
Chen
Yue
Y
Xu
Ling
L
Zhang
Qing-Hong
QH
Xia
Lu
L
Chen
Ming
M
Fu
Jun
J
D000818:Animals; D001853:Bone Marrow; D001854:Bone Marrow Cells; D001842:Bone and Bones; D002117:Calcitriol; D002478:Cells, Cultured; D003907:Dexamethasone; D010010:Osteoclasts; D051381:Rats; D017207:Rats, Sprague-Dawley; D014357:Trypsin
D003907:Dexamethasone; D014357:Trypsin; D002117:Calcitriol