Layered co-culture method for intramuscular fat cells and skeletal muscle cells of mammal

Abstract

The invention discloses a layered co-culture method for intramuscular fat cells and skeletal muscle cells of a mammal. The layered co-culture method comprises the steps: after washing and cutting a longissimus tissue on the back of a butchered animal into pieces, digesting the longissimus tissue by using type-I collagenase, and sieving a digested mixed solution; collecting filtrate, centrifuging, respectively collecting mature fat cell sap on the upper layer and muscle cells on the lower layer, respectively adding a proper amount of serum-free medium to dilute and wash, and centrifuging; adding the centrifugate on the upper layer and the muscle cell sap on the lower layer into a culture flask with the capacity of 25cm<2>, and culturing in a culture box with the temperature of 37 DEG C and the CO2 content of 5%. The mature fat cells can float and adhere to a ceiling surface on the upper layer because of small buoyancy, while muscle cells sink to the bottom of the culture flask to grow. The cell culture model can be used for layering and co-culturing the intramuscular fat cells and the muscle cells of the same muscle tissue of an animal in the same culture solution, so that a novel in-vitro cell research model is provided for researching the influence of mature fat cell secreta to muscle cell differentiation or the interaction between the muscle cells and the fat cells.

Description

technical field [0001] The invention belongs to the technical field of cell biology, and relates to a layered co-cultivation method of mammalian intramuscular adipocytes and skeletal muscle cells. Background technique [0002] The proportion, structure and distribution of animal carcass muscle and fat tissue are the main factors that determine the production performance and meat quality of meat animals. In general, muscle tissue is mainly composed of skeletal muscle cells. Adipose tissue can be divided into subcutaneous fat, visceral fat and intramuscular fat. Fat deposition in different parts mainly depends on the proliferation, differentiation and lipid maturation of preadipocytes. Among them, intramuscular fat is a beneficial fat, which is the material basis for the formation of marbled meat, directly participates in the formation of meat tenderness, juiciness and meat flavor, and is an important factor affecting the flavor of meat. In the past 20 years, scholars at hom...

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to solve the problems of difficult separation of mammalian intramuscular preadipocytes and many miscellaneous cells, and at the same time solve the problem of independent and co-culture of two kinds of cells, and provide a kind of mammalian intramuscular adipocyte and skeletal muscle Cell layered co-culture method, providing a cell model for the study of cell-cell interactions

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