Isolation and in vitro culture of intramuscular pre-adipocytes from Luxi adult Yellow cattle

Abstract

Three Luxi adult yellow steers were used to isolate and culture intramuscular preadipocyte in vitro and examine factors influencing their proliferation and differentiation. The intramuscular preadipocyte cells were taken from adipose tissues within muscles between sixth and seventh rib and cultured after digestion by collagenase Ⅰ. The results showed that the separated cells were highly homogeneous, proliferative and double timed within 62 h. When the confluent preadipocytes were treated with differentiation medium, small lipid droplets began to appear on day 2 and the number of lipid droplets rapidly increased around the nuclei on day 6. Their dynamic morphological changes, growth curve, oil O staining, and reaction to insulin and dexamethasone all verified their preadipocyte identity. Under controlled conditions, the intramuscular preadipocytes replayed their proliferation and differentiation process in vitro, especially, the proportion of cultured diploid preadipocytes reached to more than 90% after 6 run of repeated cultures were performed. In conclusion, the current study confirms that functionally active preadipocytes indeed present within muscles of China adult local breed cattle. The cell strains will be potentially used as a useful model for further understanding of the mechanism by which the intramuscular adipose is deposed in the tissues, and for improving the beef quality based on local breed beef cattle.