C/EBPα Induce The Trans-Differentiation Of Bovine Myoblasts

Meat quality traits include color, texture, hardness, tenderness, marble, etc. Among them, the amount of fat deposition and depositional mode are very important. Muscle fat is usually divided into intermuscular fat and intramuscular fat (IMF), The IMF is closely related with tenderness, flavor of meat and other food value. In recent years, Improvement IMF content to improve the quality of meat by means of genetic mesures is a beef cattle breeding’s research hotspot.The Myoblast in skeletal muscle tissue is a precursor cell, and play an important role in the growth and development of skeletal muscle. C/EBPa gene is a C/EBP family’s subtype and closely related to the proliferation and differentiation in fat cells.In order to investigate the fuctions of C/EBPa gene in the process of bovine myoblast trandifferntiating into fat, we imported the Ad-C/EBPa recombinant adenovirus in bovine myoblasts.The main results were as follows:1. Through using the IV type collagenase digestion process and differential adherence method, we cultured the bovine myoblasts, and then identified them through morphological method and induced differentiation method.2. Our study found that insulin could induce bovine myoblasts transdifferentiate into adipocyte successfully. DMEM+20%FBS+10%HS medium+insulin group’s adipogenic effect was better than DMEM+10%FBS medium+insulin group.Real-time quantitative PCR detected that during the induced transdifferentiation process, C/EBPβ, C/EBPδ and PPARy’s relative expression increased, but the C/EBPa reduced.The Myf5’s relative expression was reduced, but MyoD increased.3. Ad-C/EBPa recombinant adenovirus infection efficiency in cattle myoblasts could reach to90%, and could induce bovine myoblasts transdifferentiate into adipocyte successfully. Real-time quantitative PCR detected that during the induced transdifferentiation process, C/EBPa gene was expressed at high levels in the1st,3rd,5th and7th days,and lipid droplets increased significantly.LPL and PPARy relative expression increased in the late. Myf5relative expression decreased, MyoD increased, and Pax7increased after decresed.In summary, we successfully isolated, cultured and identified the cattle myoblasts, and established a system suitable for cattle myoblasts cultured in vitro. And then, we successfully induced bovine myoblasts differentiating into fat with using insulin and Ad-C/EBPα recombinant adenovirus, providing evidence about the role of C/EBPα gene in bovine adipose tissue during growth and development, and lay a theoretical basis for further study of the molecular mechanisms of bovine fat metabolism and meat quality improvement.