| Objective To investigate the effects of different culture medium on the differentiation of RAW264.7 cells into osteoclasts, in order to optimize the culture conditions of osteoclasts differentiation. Methods The experiment was divided into three groups: high glucose DMEM differentiation medium group (DMEM group), high glucose DMEM/α-MEM differentiation medium group (DMEM/α-MEM group), α-MEM differentiation medium group (α-MEM group). Tartrate-resistant acid phosphatase(TRAP) staining was used to observe the formation of mature osteoclasts in each group. Toluidine blue staining was used to quantitatively analyze the area of bone lacuna to observe the function of osteoclasts. Real time PCR was used to detect the expression of NFATc-1, c-Fos and TRAF-6 mRNA. Results Typical TRAP+ osteoclasts were observed in all groups. Compared with DMEM group, the number of TRAP+ osteoclasts in DMEM/α-MEM group and α-MEM group was significantly increased (P < 0.01). However, the morphology of every group are slightly different. Compared with DMEM group, the mRNA expressions of NFATc-1, c-Fos and TRAF-6 in α-MEM group were significantly increased (P<0.01, P<0.05); Compared with the DMEM group, the mRNA expression of NFATc-1 in DMEM/α-MEM group was significantly increased (P<0.01), and the mRNA expression of c-Fos and TRAF-6 had a tendency to increase, but there was no statistical difference, there was no significant different between α-MEM group and DMEM/α-MEM group. Conclusion High glucose DMEM differentiation medium, high glucose DMEM/α-MEM differentiation medium and α-MEM differentiation medium can induce RAW264.7 cells to differentiate into osteoclasts, but from the number, morphology and function of osteoclasts, α-MEM medium is more suitable for osteoclast differentiation. |