成年鲁西牛肌内前脂肪细胞的分离培养  

Isolation and In vitro Culture of Intramuscular Preadipocyte from Luxi Adult Yellow Cattle

作　　者：万荣[1] 丁健[1] 周振明[1] 任丽萍[1] 孟庆翔[1] 

机构地区：[1]中国农业大学动物营养学国家重点实验室,动物科技学院/肉牛研究中心,北京100094

出　　处：《农业生物技术学报》2007年第3期419-423,共5页Journal of Agricultural Biotechnology

基　　金：农业部948项目(No.2006-G47)资助

摘　　要：选取来自3头成年育肥鲁西黄牛阉牛第6～7肋间的肌肉内脂肪组织,利用Ⅰ型胶原酶消化法分离获得牛肌内前脂肪细胞。培养出的细胞成分均一,增殖旺盛,倍增时间为62h;体外诱导分化2d时,培养细胞胞质内开始有小脂滴出现,而至第6天时,细胞胞质内小脂滴明显增多,并且有少数开始融合成大脂滴。经形态学动态变化观察、生长曲线、油红O脂肪染色提取法及对胰岛素和地塞米松反应的测定,证明是功能活跃的前脂肪细胞,并在体外重现了其增殖分化过程,同时经染色体核型分析分离获得的牛肌内前脂肪细胞经6次传代培养后,其二倍体细胞数超过90%,基本都保留着正常细胞的基本遗传特征。本实验证明,成年鲁西黄牛肌肉内群确实存在前脂肪细胞。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.

关 键 词：鲁西黄牛 肌内前脂肪细胞 体外培养 大理石纹 

分 类 号：S188[农业科学—农业基础科学]