畜牧兽医学报 ›› 2012, Vol. 43 ›› Issue (9): 1346-1352.doi:

• 遗传繁育 • 上一篇    下一篇

莱芜猪前体脂肪细胞的原代培养及诱导分化研究

胡艳霞,曾勇庆*,崔志峰,杨伦,陈伟,陈其美,石元,祝洪磊   

  1. (山东农业大学动物科技学院,泰安 271018)
  • 收稿日期:2011-12-21 出版日期:2012-09-25 发布日期:2012-09-25
  • 通讯作者: 曾勇庆,E-mail: yqzeng@sdau.edu.cn
  • 作者简介:胡艳霞(1986-),女,河南开封人,硕士生,主要从事动物遗传育种学研究,E-mail: huyanxia1986@126.com
  • 基金资助:

    国家转基因重大专项(2011ZX08006-002);国家高技术研究发展计划“863”重点项目(2008AA101008);山东省现代农业(生猪)产业技术体系建设专项;山东省农业良种工程重大课题(2009LZ012-03)

Primary Culture and Induced Differentiation of Laiwu Black Pig Preadipocytes

HU Yan-xia, ZENG Yong-qing*, CUI Zhi-feng, YANG Lun, CHEN Wei, CHEN Qi-mei,
SHI Yuan, ZHU Hong-lei   

  1. (College of Animal Science and Technology, Shandong Agricultural University, Taian
    271018, China)
  • Received:2011-12-21 Online:2012-09-25 Published:2012-09-25

摘要: 为了建立莱芜猪前体脂肪细胞的体外培养体系,探索猪脂肪组织增生的生物学特征。本研究采集1日龄莱芜猪仔猪颈、背部皮下脂肪组织,采用胶原酶与胰蛋白酶相结合的方法,分离原代前体脂肪细胞,进行前体脂肪细胞的原代和传代培养,以及前体脂肪细胞诱导分化的研究,并采用Real-time PCR方法检测前体脂肪细胞分化过程中关键基因PPARγ的表达。结果表明,分离的猪原代前体脂肪细胞约8 h后开始贴壁,贴壁的细胞呈短梭形或不规则的三角形,第3天多数细胞进入指数生长期,呈成纤维细胞样形态,细胞生长曲线近似S形;诱导培养结果显示,少量细胞在第2天开始出现脂滴,大多数细胞在第3天出现脂滴,第7~8天小脂滴逐渐融合成大脂滴,在第12天左右融合成大脂滴,油红O染色呈橘红色。在前体脂肪细胞分化过程中PPARγ基因mRNA的表达量逐渐增加,在诱导后的48 h表达量最高,随后其表达丰度逐渐下降。本试验成功建立了莱芜猪前体脂肪细胞培养体系和体外诱导分化模型,为进一步研究莱芜猪前体脂肪细胞增殖与分化机制和猪体脂肪的沉积奠定基础。

Abstract: The study aimed to establish the primary culture system of Laiwu Black pig preadipocyte for better studying the properties of hyperplasia of Laiwu Black pig adipose tissue. Preadipocytes were isolated from subcutaneous adipose tissue of 1 day piglet by the combination of collagenase and trypsin digestion and were successfully cultured. The induction and differentiation of preadipocytes were studied, the expression of the important transcription factor PPARγ in the process of preadipocytes differentiation were detected. The result showed that the isolated cells started adherence at 8 h after initiation of primary culture and displayed short spindle or irregular triangle. On 3rd day, the cells entered logarithmic growth phase, displayed fibroblast-like, and the cells growth showed ‘S’ shape; the induction result showed that fat droplets occurred in small amount of cells on 2nd day, in large amount on 3rd day, those small fat droplets gradually converged into big ones from 7th to 8th day, and on the 12th day become to one big fat droplets, the adipocytes showed jacinth by oil red O staining; the PPARγ mRNA levels gradually increased during the process of the preadipocytes differentiation, and reached the top at 48 h after induced differentiation culture, then the PPARγ mRNA levels gradually decreased. The Laiwu Black pig primary culture system and the model of the induced differentiation in vitro were established successfully. This study laid the basis for further researching Laiwu Black pig adipocyte proliferation, differentiation and porcine fat deposition.

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