| 摘要: |
| 【目的】建立牛前体脂肪细胞的体外分离培养方法，研究牛前体脂肪细胞的生物学特性和分化特征，为研究牛脂肪发育和脂肪沉积的机制提供一种简便有效的细胞模型。【方法】采用Ⅰ型胶原酶消化法自新生牛脂肪组织中获得前体脂肪细胞，对培养的牛前体脂肪细胞进行形态学观察、生长曲线测定、油红O染色及脂肪细胞标志基因LPL、PPAR-r 和脂联素表达研究。【结果】80%的牛前体脂肪细胞接种后12 h开始贴壁，4 d后开始向脂肪细胞转化，10 d后绝大部分细胞脂滴融合，细胞脂肪含量增加；分离的牛前体脂肪细胞接种后1～2 d生长缓慢，3～8 d进入对数生长期，9 d后进入平台期，之后细胞数目开始下降。经胰岛素诱导分化过程中，LPL在牛前体脂肪细胞分化的早期开始表达，PPAR-γ在分化的中期开始高度表达，而脂联素在牛前体脂肪细胞分化的前期未检测到，在细胞分化后期高度表达。【结论】用Ⅰ型胶原酶消化法可获得大量的牛前体脂肪细胞。培养的牛前体脂肪细胞成分均一、生长旺盛，经胰岛素诱导后能稳定的向脂肪细胞分化。 |
| 关键词: 牛 前体脂肪细胞 生物学特性 诱导分化 分离培养 |
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| 基金项目:国家肉牛牦牛产业技术体系项目(CARS-38)；国家自然科学基金项目(31272411)；国家“863”计划项目(2011AA100307-02，2013AA102505)；国家科技支撑计划项目(2011BAD28B04 03，2012BAD12B07)；教育部长江学者和创新团队发展计划项目(IRT0940)；农业部科研杰出人才及其创新团队发展计划项目（2012-64-52）；陕西省科技统筹创新工程计划项目（2011KTCL02-07） |
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| Primary culture and differentiation of bovine preadipocytes |
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GUO Hong-fang,ZAN Lin-sen,SUN Yong-gang
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| Abstract: |
| 【Objective】The study was to explore and establish a stable culturing system for bovine preadipocytes in vitro and study the biological and differentiation characteristics of bovine preadipocytes.【Method】Preadipocytes were isolated from adipose tissue of newborn bovine by type Ⅰ collagenase.Cultured preadipocytes were identified by morphological change,growth curve,oil red O staining,and mRNA expression of lipoprotein lipase gene (LPL),peroxisome proliferators activated receptor-γ gene (PPAR-r) and adiponectin gene.【Result】80% bovine preadipocytes began to paste the wall after inoculation for 12 h,and the cells began to transfer to fat cells after 4 days.10 days later,lipid droplets of most cells fused,and fat contents increased.Bovine preadipocytes grew well in vitro.On days 1-2,cells grew slowly,and the cells entered logarithmic growth phase during days 3-8.The growth of cells became stable from day 9,since when the number of cells decreased.During the differentiation by insulin,LPL gene expressed in the early stage and PPAR-γ gene expressed in the middle stage.The expression of adiponectin gene was not detected in the early stage,but it was highly expressed in mature adipocytes.【Conclusion】Preadipocytes isolated from adipose tissue of newborn bovine by type Ⅰ collagenase grew well and could be differentiated into mature adipocyte in vitro. |
| Key words: bovine preadipocyte biological characteristics differentiation primary culture |
