. 2010, 28(1001): 44-47.
This paper studies the identification method by inducing RAW264.7 into osteoclast, to find the best procedure, for in vitro functional researches, as a preferable preosteoclast model in respect of its osteoclast characteristic gene expression profile. The inoculate RAW264.7 in 96 well plates, cultured with the bone ground section, of a concentration of 100ng/mL, was used to induce mouse sRANKL for 10 days, which was then examined with respect to morphology and functional identification and detection, using HE staining, Toluidine Blue staining, TRAP staining and SEM observation. It is concluded that with the increase of the induced time, the number of polykaryocyte cells and the amount of Howship's lacuna increase. Among all kinds of methods of identification, HE staining and Toluidine Blue staining are simple but not reliable; the SEM observation is accurate , but complex; and TRAP staining is accurate and shows strong specificity, but is expensive. The results show that the TRAP staining is the best method in view of morphology identification, SEM observation is the best method in view of functional identification. RANKL can induce RAW264.7 cell into mature osteoclast for morphology and functional identification and detection, which can serve as a basis for our future experimental research about mature osteoclast.
