Molecular Nutrition

Isolation and Purification of Intramuscular Lipid Droplets in Porcine Skeletal Muscle

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  • 1. Tianjin Institute of Animal Husbandry and Veterinary Medicine, Tianjin Academy of Agricultural Sciences, Tianjin 300381, China;
    2. Institute of Chemical Technology, Tianjin University, Tianjin 300072, China;
    3. College of Animal Science and Technology, Northwest A & F University, Yangling 712100, China;
    4. Institute of Animal Sciences of Chinese Academy of Agricultural Sciences State Key Laboratory of Animal Nutrition, Beijing 100193, China

Received date: 2018-06-26

  Online published: 2019-01-16

Abstract

This experiment aimed to explore the establishment of an optimal method for the isolation and purification of intramuscular lipid droplets from porcine skeletal muscle, which would lay the foundation for further revealing the regulation mechanism of intramuscular fat formation. Longissimus dorsi muscle tissue from body weight of (120±2) kg "Duroc×Landrace×Yorkshire" ternary hybrid finishing pigs was obtained after slaughter. The isolation of intramuscular lipid droplets of longissimus dorsi muscle was performed by ultracentrifugation. Explore the influence of different homogenization methods, temperature, buffer pH and phenylmethanesulfonyl fluoride (PMSF) on the isolation efficiency, and further determine the optimal centrifugal force and centrifugation time to purify lipid droplets. The results showed that the skeletal muscle was homogenized by a fully automatic tissue grinder, and the intramuscular lipid droplets were obtained by adding PMSF in 4℃, pH=7.4 buffer. With the purification conditions at 4℃, 230 000×g centrifugation with 30 min, the lipid droplets were the most abundant, the structure was complete, and the purity was the highest. In conclusion, the optimized isolation and purification method can obtain intramuscular lipid droplets of porcine skeletal muscles with complete morphology and high purity, which can open up new avenues to study intramuscular fat generation and the molecular regulation mechanism.

Cite this article

YAN Jun, MA Yanxuan, MU Shuqin, TANG Xiangfang, ZHENG Zi, LI Ning, LIU Chenlong, LI Qianjun, SUN Chao . Isolation and Purification of Intramuscular Lipid Droplets in Porcine Skeletal Muscle[J]. Chinese Journal of Animal Nutrition, 2019 , 31(1) : 266 -273 . DOI: 10.3969/j.issn.1006-267x.2019.01.033

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