Quantitative evaluation of bone resorption activity of osteoclast-like cells by measuring calcium phosphate resorbing area using incubator-facilitated and video-enhanced microscopy.
Quantitative evaluation of the ability of bone resorption activity in live osteoclast-like cells (OCLs) has not yet been reported on. In this study, we observed the sequential morphological change of OCLs and measured the resorbing calcium phosphate (CP) area made by OCLs alone and with the addition of elcatonin utilizing incubator facilitated video-enhanced microscopy. OCLs, which were obtained from a coculture of ddy-mouse osteoblastic cells and bone marrow cells, were cultured on CP-coated quartz cover slips. The CP-free area increased constantly in the OCLs alone, whereas it did not increase after the addition of elcatonin. This study showed that analysis of the resorbed areas under the OCL body using this method enables the sequential quantitative evaluation of the bone resorption activity and the effect of several therapeutic agents on bone resorption in vitro.
Morimoto
Yoshitaka
Y
Department of Orthopaedic Surgery, Hamamatsu University School of Medicine, 1-20-1 Handayama, Higashi-ward, Hamamatsu 431-3192, Japan. morimo2@nifty.com
Hoshino
Hironobu
H
Sakurai
Takashi
T
Terakawa
Susumu
S
Nagano
Akira
A
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000502:physiology*; D001862:Bone Resorption; D002116:Calcitonin / Q000031:analogs & derivatives / Q000378:metabolism; D002130:Calcium Phosphates / Q000378:metabolism*; D018920:Coculture Techniques / Q000295:instrumentation / Q000379:methods; D008297:Male; D051379:Mice; D008858:Microscopy, Phase-Contrast; D018715:Microscopy, Video / Q000379:methods; D010010:Osteoclasts / Q000166:cytology / Q000502:physiology*
D002130:Calcium Phosphates; D002116:Calcitonin; C020243:calcium phosphate; C012755:elcatonin