Interaction between mouse macrophages RAW264.7 and myoblasts C2C12 in a co-culture system

Di-san junyi daxue xuebao

LUO We¹  LI Xian¹  WANG Bofa¹  ZHOU Yue¹  AI Lei³ 

[1] Department of Exercise Physiology, Beijing Sport University, Beijing, 100084;Department of Sports Rehabilitation, Nanjing Sport Institute, Nanjing, Jiangsu Province, 210014;Second Center of Competitive Sports Science Research, Jiangsu Provincial Research Institute of Sports Science, Nanjing, Jiangsu Province, 210033, China;

关键词: myoblasts;    macrophages;    co-culture;    myogenic differentiation;    polarization phenotype;   

DOI  :  10.16016/j.1000-5404.201905173

来源: DOAJ

【 摘 要 】

Objective To establish a co-culture system to observe the interactions between mouse macrophage RAW264.7 and myoblast C2C12 cells. Methods RAW264.7 and C2C12 cell lines were co-cultured in a Transwell chamber containing conditioned medium for myogenic differentiation, and the changes in the cell morphology were observed under a phase-contrast microscope. On days 1, 3 and 5 of co-culture, the cell proliferation was assessed with CCK-8 assay, and the protein expressions of Myf5, MyoD, myogenin, iNOS and Arg-1 were detected using Western blotting; the concentrations of interleukin-1β (IL-1β) and IL-10 in the supernatant were detected using enzyme-linked immunosorbent assay (ELISA). Results Co-culture with RAW264.7 cells obviously accelerated the myogenic differentiation and promoted the formation of multinucleated myotubes in C2C12 cells. Compared with the cells cultured alone, C2C12 cells in the co-culture system showed significantly inhibited protein expression of Myf5 (P < 0.05) and enhanced expressions of MyoD (P < 0.05) and myogenin (P < 0.01) on day 1, and inhibited expression of Myf5 (P < 0.01) and increased expression of MyoD (P < 0.01) on day 3; significantly lowered cell viability (P < 0.05) and Myf5 expression (P < 0.01) with increased area of myotubes (P < 0.01) were observed on day 5. The co-culture did not produce significant effect on the morphology of RAW264.7 cells, but caused significant inhibition of iNOS expression (P < 0.01) and IL-1β secretion (P < 0.05) and increased Arg-1 expression (P < 0.01) on day 1 of the co-culture; significantly lowered iNOS expression and IL-1β secretion (P < 0.01) and increased Arg-1 expression (P < 0.01) and IL-1β secretion (P < 0.01) occurred in RAW264.7 cells on day 3 of the co-culture, and on day 5, the cells showed more obvious clustering with significantly enhanced proliferation (P < 0.05), lowered iNOS expression and IL-1β secretion (P < 0.01), and increased secretion of IL-10 (P < 0.05). Conclusion Co-culturing C2C12 cells with RAW264.7 cells accelerates the myogenic differentiation of the former and promotes the proliferation and M2 polarization of the latter.

【 授权许可】

Unknown