绵羊肌内脂肪前体细胞的体外培养及分选

为建立绵羊肌内脂肪前体细胞的体外培养模型,选用组织块法培养绵羊肌内脂肪前体细胞,制备用荧光染料PE(Phycoerythrin)标记的兔抗绵羊Pref-1蛋白的多克隆抗体,通过流式细胞术分离纯化脂肪前体细胞,对纯化后的细胞进行诱导分化以及油红O染色鉴定。结果表明:利用流式细胞仪能分选纯化出被标记的肌内脂肪前体细胞且分选率近30%;纯化后得到的细胞是功能活跃的脂肪前体细胞。研究表明,用绵羊羔羊肌肉组织采用组织块培养法可以建立绵羊肌内脂肪前体细胞培养模型,抗绵羊Pref-1蛋白多克隆抗体可以作为纯化脂肪前体细胞的工具。

In vitro culture and purification of ovine intramuscular preadipocyte

This study was conducted to set up a method of ovine intramuscular preadipocyte culture in vitro.First, ovine intramuscular preadipocytes were isolated and cultured in vitro from the longissimus dorsi muscle tissues of a one-month-old lamb.During culture, the dynamic morphological changes of cultured cells were conducted.Then the synthetic peptide fragments of ovine Pref-1 were used to immunize the rabbit to obtain rabbit anti-sheep Pref-1 polyclonal antibody which was labeled with phycoerythrin (PE).And then the cells were separated and purified by the flow cytometer.The reaction of purified preadipocytes to insulin and dexamethasone was observed, and identified with oil red O.The result showed that lamb muscle tissues could be used to culture intramuscular preadipocytes in vitro.The specificity of the antibody was sharp.And the ratio of purification was up to 30%.In conclusion, the study confirms that muscle tissues of lamb could be used to establish a culture model of intramuscular preadipocytes in vitro, and rabbit anti-sheep Pref-1 polyclonal antibody could be a tool to purify preadipocytes.