Study on differentiation of muscle derived stem cells in the rat penile corpus cavernosum into adipose cells

Abstract

Objective We observed the potential of the rat penile corpus cavernosum muscle-derived stem cells (MDSCs) differentiating into adipose cells in the presence of adipose differentiation culture medium in vitro.Methods MDSCs were purified through density gradient centrifugation and differential attachment.PP6 cells were obtained and detected expression of stem cell markers Sca-1 by immunofluorescence.The third passages PP6 cells were induced to differentiate to adipose cells within 21 days of adipose differentiation culture medium stimulation,while the control group used stem cells culture medium.Induced cells was detected by oil red O staining at 0,7,14 and 21 d respectively in order to observed whether there was a lipid droplets.Results PP6 cells expressed stem cell markers Sca-1 and PP1 cells hardly expressed Sca-1.On the first three days,we observed that the shape of cells had changed from long spindle-shape to circular or triangular,closely arranging as imbricate,meanwhile nucleus clarity and proliferation ability of cells had significantly slowed down.Both of groups had not changed by oil red O staining.On the seventh day,we observed that a small amount of high refractive small lipid droplets was in cytoplasm under phase contrast microscope,uniform size,which mainly concentrated around the cells nucleus,and was dyed red by oil red O staining.On the fourteenth day,the number of lipid droplets in cells was increased,lipid droplets blended into large,and the cells nucleus were crowded to one side or disappeared.On the twentieth day,cell differentiation,lipid droplets volume and number in cytoplasm reached to peak,and even continued to induce,but lipid droplets did not increased.In control group,however,cell morphology did not change and oil red O staining was negative.Conclusion Rat corpus cavernosum MDSCs have potential to differentiate into adipose cells,and MDSCs multi-directional differentiation potential is further confirmed. Key words: Muscle-derived stem cells; Differentiation; Adipose cells; Oil red O