猪肌源性前体脂肪细胞的分离培养和成脂诱导分化研究

doi:10.11843/j.issn.0366-6964.2018.12.010

Abstract

Abstract:

The aim of this study was to establish the culture system of porcine myogenic adipocytes cell in vitro, and to understand the mRNA expression regulation of adipogenesis related genes during adipogenic differentiation process and to provide experimental materials for further studying on the mechanism of intramuscular fat deposition in pig. Samples of longissimus dorsi muscle of the 15 days old piglets were collected and the precursor adipocytes cell was obtained by digested with type Ⅱ collagenase, and the cell were separated, primary and passage cultured and adipogenic induction. The morphology of cell were observed and the growth curve was determined. The adipogenesis was measured by Oil red O staining and the mRNA expression of adipogenesis related genes, such as PPARγ, FABP4, C/EBPα, C/EBPβ, ZFP423, SREBP1, PRMD16 and HSL were detected by qRT-PCR during induced differentiation. The results showed that a small number of myogenic preadipocytes cells began to adhere to the cell wall at about 6-7 h, and the shape of the cells were round, small shuttle and irregular, and became uniform in fibroblast-like appearance after cell passage. The growth curve was typical "S" shaped. All genes detected in this study were expressed at early stage of adipogenic differentiation after adding differentiation medium, in which the mRNA expression of FABP4, C/EBPα, SREBP1 and PPARγ at early stage was very significantly higher (P<0.01) and then was decreased significantly (P<0.01). There was no significant change for the mRNA expression of C/EBPβ,ZFP423 and PRMD16 genes during different differentiation stages. The mRNA expression of HSL was significantly reduced with the differentiation process (P<0.01). Oil red O staining was red after induction for 12 days. In conclusion,The system of the isolation, culture and adipogenic differentiation of porcine myogenic adipocytes cell was successfully established, and the expression and regulation of key genes involved in adipogenesis was investigated during cell differentiation process, which provide the foundation for further studying on the mechanism of intramuscular fat deposition and improvement of meat quality in pig production.

CLC Number:

S828.2

ZHANG Ning-fang, CHENG Zhi-min, LE Bao-yu, QIN Ben-yuan, WANG Yuan-yuan, WANG He-jie, GAO Peng-fei, GUO Xiao-hong, LI Bu-gao, CAO Guo-qing. Isolation, Culture and Adipogenic Differention of Pig Myogenic Preadipocytes Cell[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(12): 2612-2621.

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Isolation, Culture and Adipogenic Differention of Pig Myogenic Preadipocytes Cell

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