Abstract:

Objective To compare the three methods on the effect of myogenic differentiation of porcine dedifferentiated adipocytes (DA), in order to screen the better method for inducing myogenic differentiation of porcine DA. Methods The glucocorticoid, galectin-1 and 5-azacytidine(5-aza) were used to induce myogenic differentiation of porcine DA for 6 days, 15 days and 21 days respectively, and then the morphology of cell differentiation was observed by inverted microscope respectively. After 21 days of induction, the expression of muscle specific protein which desmin and myosin heavy chain(MyHC) was detected by indirect immunofluorescence, and the expression of myogenin(MyoG) mRNA was detected by Real-time PCR. Each method was used for detecting induction effect in 3 groups and repeat three times in each group. Results The morphological observation of induced cell showed that the best myotube morphology and the largest number of multinucleated cells was found via galectin-1 induced, followed by 5-aza, however the higher percent of cell death was occurred during 5-aza induction. The irregular cell morphology, multiform radial protrusion, and some cells of adipogenic differentiation were observed by glucocorticoid induction. The indirect immunofluorescence showed that the strong positive of Desmin and MyHC as well as a high expression of MyoG mRNA was observed in induced cells via galectin-1 method and glucocorticoid method, while the weak positive expression of these two kinds of protein with a low MyoG mRNA level was found in induced cells by 5-aza induction. Conclusion Judged from myotube morphology, differentiation of purity and viability, specific protein and gene expression, we suggest that galectin-1 method is superior to the glucocorticoid and 5-aza method for inducing myogenic differentiation of porcine DA.

Key words: Dedifferentiated adipocyte, Myogenic differentiation, Glucocorticoid, Galectin-1, 5-azacytidine, Immunofluorescence, Porcine