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. Author manuscript; available in PMC: 2020 Aug 1.
Published in final edited form as: J Cell Biochem. 2019 Feb 25;120(8):12450–12460. doi: 10.1002/jcb.28512

Figure 1. M-CSF pretreatment decreases mouse bone marrow cells OC differentiation.

Figure 1.

A) A schematic diagram of the OC differentiation protocol using mouse bone marrow cells (BMC). BMC were isolated from mouse femurs, red blood cells lysed and the total population adhered to plastic cell culture dishes for 14-18h (1adBMC). After 18 h the non-adherent cells were removed and replated onto plastic cell culture dishes (2adBMC). After three days, 1adBMC and 2adBMC were treated with or without M-CSF for 3 days, followed by addition of OC differentiation medium (M-CSF and RANKL).

B) TRAP staining shows that plastic adBMC can be effectively induced into multinuclear TRAP+ OC with M-CSF and RANKL, particularly the slow adherent cells (2adBMC) in the absence of M-CSF pretreatment. C). M-CSF pretreatment markedly decreased OC formation of 1adBMC and 2adBMC. D). Dose response of M-CSF pretreatment shows that pretreatment with as little as 5 ng/ml M-CSF was sufficient to inhibit OC formation of 2adBMC. E). Representative images of TRAP stained OC from a time course analysis of M-CSF pre-treatment shows M-CSF inhibited OC differentiation of 1adBMC. F). Quantification from time course analysis in E. G). Statistical analysis shows that M-CSF pre-treatment resulted in higher variation in the experiments from E. Note: **: p<0.01; ***: p<0.001.