摘要
为建立松辽黑猪前体脂肪细胞分化体系,了解其增殖规律及相关的脂肪标志基因在细胞分化过程中的相对表达。选用7日龄松辽黑猪,用Ⅰ型胶原酶消化脂肪组织得到前体脂肪细胞,传代后进行形态学观察并绘制生长曲线,利用传统"鸡尾酒"法进行诱导分化,油红O对分化后的细胞进行染色,判断分化效果,提取总RNA,对成脂标志基因进行相对表达量分析。结果表明:分离获得的松辽黑猪原代前体脂肪细胞可贴壁生长,形似成纤维细胞,细胞生长曲线符合细胞生长规律,近似"S"形;诱导分化后的细胞具有成熟脂肪细胞的典型特征,细胞内聚集大量脂滴;诱导后第10天,细胞内甘油三酯含量极显著提高;成脂标志基因PPARγ、C/EBPα在分化后的细胞中相对表达量极显著升高(P<0.01),而FoxO1在分化前相对表达量最高,第5天表达量与第0天相比极显著下降(P<0.01),但第10天的表达量极显著高于第5天(P<0.01),且第10天表达量与第0天无显著差异(P>0.05)。试验建立了松辽黑猪前体脂肪细胞的分离体系,为进一步研究松辽黑猪脂肪沉积与代谢机制以及改善肉质提供了依据。
The aim of this study was to establish Songliao black pig preadipocyte differentiation system and to understand its proliferation pattern and the relative expression of marker genes during the differentiation.Adipose tissue was collected from a seven-day-old Songliao black piglet and the preadipocytes were isolated using typeⅠcollagenase digestion method.The traditional"cocktail"induction method was used to induce cell differentiation.Adipocytes were stained by oil red O for morphological observation.The total RNA of the cells was extracted,and the relative expression level of the marker gene in the cells was detected by qRT-PCR.The results showed that the isolated primaryadipocytes of Songliao black pigs could grow adherently and resembled fibroblasts.The cell growth curve was in accordance with the cell growth pattern,which was similar to the"S"shape.The differentiated cells had typical characteristics of mature adipocytes,which accumulated large amounts of lipid droplets in the cells;The expression of PPARγand C/EBPαin the differentiated cells were highly expressed(P<0.01),While the expression of FoxO1 was the highest before differentiation.The expression of FoxO1 on the 5 th day was significantly lower than that on the 0 th day(P<0.01),but the expression on the 10 th day was significantly higher than that on the 5 th day(P<0.01),and the expression level on the 10 th day was not significantly different from the 0 th day(P>0.05).This study built the porcine preadipocyte differentiation system in vitro,which would provide materials for fat deposition and metabolism mechanism and improve meat quality in Songliao black pig.
作者
朴政玉
金一
于永生
罗晓彤
程咪咪
PIAO Zhengyu;JIN Yi;YU Yongsheng;LUO Xiaotong;CHENG Mimi(College of Agriculture,Yanbian University,Yanji 133002,China;Branch of Animal Husbandry Science,Jilin Academy of Agricultural Sciences,Gongzhuling 136100,China)
出处
《吉林农业大学学报》
CAS
CSCD
北大核心
2020年第3期337-342,共6页
Journal of Jilin Agricultural University
基金
吉林省科技厅重点科技攻关项目(20150204079NY)
关键词
松辽黑猪
前体脂肪细胞
脂肪细胞分化
Songliao black pig
preadipocyte
adipocyte differentiation
作者简介
朴政玉,女,在读博士,研究方向:动物遗传育种与繁殖;通信作者:金一;通信作者:于永生
