MyoD，Mef2c And Myf6Induced Difefrentiation Of Porcine MSCs Into Myoblast-like Cells

Coronary heart disease and other cardiovascular diseases have become theleading cause of human death in recent years. Coronary heart disease could causemyocardial infarction, which makes the activities of heart myocardial cells to beweakened, leading to ventricular reconstruction, resulting in arrhythmia heart failureor even death. The regenerative capacity of myocardial cells, which are terminallydifferentiated cells, is limited, so they could not be repaired or differentiated once bedamaged. Bone marrow mesenchymal stem cells (mesenchymal stem cells, MSCs)are somatic stem cells, which could be induced to myoblast differentiation in thecertain culture conditions. It can be concluded that MSCs have a good applicationprospect in the damaged repair tissue engineering treatment.Both of MyoD and Myf6belong to myogenic regulatory factors (MRFs), controlthe proliferation and differentiation of muscle cells. Muscle enhance factor2C (Mef2c)is an important transcription factor which could be used as a specific sign ofmyocardial differentiation. The combination treatment of MyoD and Mef2c couldinduce a large number of muscle-specific target genes expression, leading to the cellsdifferentiate to the myoblast.In this experiment we separated and cultured porcine bone marrow mesenchymalstem cells, and identified the specific surface antigen of pMSCs using the flowcytometry instrument. To test the differentiation capacity of pMSC, we usedifferentiation medium induce pMSCs to adipogenic and osteogenic. By constructingthe MyoD, Myf6and Mef2c lentiviral expression vectors and packaging lentivirus.Then, they were used to infecte pMSCs cells in vitro and successfully induced tomyoblast differentiation. The mRNA and protein level of MyoD, Myf6and Mef2c in pMSCs were upregulated. And we found that the level of MyoG and CKM were alsoraised. That Means MyoD, Myf6and Mef2c were successfully over-expressed in theMSCs. Induced MSC express important myoblast factor MyoG and CKM. It provedthat MSC cells were induced to myoblasts differentiation by overexpression of MyoD,Myf6, Mef2c using lentiviral overexpression system. The results settle thefoundation for further molecular biological mechanism of MSC differentiation tomyoblasts.