中国畜牧兽医2025，Vol.52Issue(10)：4563-4574,12.DOI:10.16431/j.cnki.1671-7236.2025.10.004
哈萨克羊胎儿期臀脂前体脂肪细胞的增殖和分化特性研究
Study on the Proliferation and Differentiation Characterization of Precursor Adipocytes from Rump Adipose in Kazakh Sheep During Fetal Period
摘要
Abstract
[Objective]The aim of this study was to isolate preadipocytes during the critical developmental phase of rump adipogenesis in Kazakh sheep,and investigate the developmental mechanism of rump adipose deposition in Kazakh sheep through histological and cytological analysis,so as to lay a foundation for investigating the molecular mechanisms underlying rump adipose deposition in sheep.[Method]The embryos of Kazakh sheep at 50(E50),60(E60),70(E70),80(E80),94(E94),and 105(E105)days were collected,and the morphological characteristics of rump fat was observed.Oil Red O staining and adipogenic differentiation gene expression analysis were performed on rump adipose from embryos in Kazakh sheep during E70,E80,and E94 periods to determine the crucial phases of rump adipose deposition.Preadipocytes of rump adipose in Kazakh sheep during E70,E80,and E94 periods were isolated using a type Ⅰcollagenase stepwise digestion method.All three stages cells were subjected to 6 days continuous culture for growth curve plotting.Meanwhile,the proliferative capacity of preadipocytes from three embryonic stages was compared.Adipogenic induction and differentiation were performed on rump adipose preadipocytes in Kazakh sheep during E70,E80,and E94 periods.The differentiation potential was evaluated through Oil Red O staining,lipid droplet content quantification,and Real-time quantitative PCR analysis of adipogenic differentiation gene expression,and a comparative analysis of the adipogenic differentiation characteristics of preadipocytes across different embryonic stages was conducted.[Result]There was obvious rump adipose in Kazakh sheep in E70 period,with visible lipid droplets initially forming rump at E80 period,followed by significant accumulation of lipid droplets in E94 period.Additionally,the expression of genes related to adipogenic differentiation gradually increased with gestation time.Notably,there was a fold increase from E80 to E94 periods.Furthermore,the precursor adipocytes at three developmental periods(E70,E80,and E94)from rump adipose isolated by type Ⅰ collagenase stepwise digestion method exhibited fibroblast-like with robust growth activity.Preadipocytes from all three periods were treated with induced differentiation to produce mature lipid droplets.The expression of adipogenic differentiation genes(CEBPα,PPARγ,LPL,and FABP4)in post-induction were significantly higher than that in pre-induction(P＜0.05).Compared with E70 or E94 periods,preadipocytes in E80 period had a higher capacity for lipid differentiation,earlier lipid droplet formation(3 day),and higher droplet content.[Conclusion]The proliferation of rump adipose preadipocytes from isolated Kazakh sheep embryos during E70,E80 and E94 periods was well active.The cells could be induced to differentiate to form mature lipid droplets,and the differentiation ability of rump adipose preadipocyte was stronger in E80 period than that in the other two periods.关键词
哈萨克羊/胎儿期/臀脂/前体脂肪细胞/原代分离培养/成脂分化相关基因Key words
Kazakh sheep/fetal period/rump adipose/precursor adipocytes/primary isolation and culture/lipidogenic differentiation-related genes分类
农业科技引用本文复制引用
张倩雯,李婧平,岳成广,王嘉俊,李忠慧,李文蓉..哈萨克羊胎儿期臀脂前体脂肪细胞的增殖和分化特性研究[J].中国畜牧兽医,2025,52(10):4563-4574,12.基金项目
新疆维吾尔自治区重点实验室开放课题"永生化绵羊前体脂肪细胞系的建立及应用研究"(2023D04034) （2023D04034）
国家重点研发计划课题"绵羊高繁殖力及优异毛用种质资源精准鉴定"(2021YFD1200901) （2021YFD1200901）
