A method for isolating intramuscular fat cells of Qinchuan beef cattle

A technology of intramuscular fat cells and separation method, which is applied in the field of separation of intramuscular fat cells of Qinchuan beef cattle, can solve the problem of unclear definition of the mechanism of preferential accumulation of intramuscular fat, and achieve the purpose of increasing subcutaneous and visceral fat deposition and improving muscle fat accumulation. Inner fat deposition and the effect of increasing the ratio of positive cells

Active Publication Date: 2022-05-27
NORTHWEST A & F UNIV
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Problems solved by technology

However, the mechanisms leading to the preferential accumulation of intramuscular fat remain poorly defined so far

Method used

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  • A method for isolating intramuscular fat cells of Qinchuan beef cattle
  • A method for isolating intramuscular fat cells of Qinchuan beef cattle
  • A method for isolating intramuscular fat cells of Qinchuan beef cattle

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Embodiment Construction

[0025] During the research process, the applicant found that more positive preadipocytes could be obtained by digesting the longissimus dorsi muscle of Qinchuan beef calf with type II collagenase, compared with other methods such as type I collagenase digestion, or with adult cattle. There are some differences in intramuscular adipocytes obtained from the longissimus dorsi samples.

[0026] The applicant found that the longissimus dorsi muscle tissue of newborn cattle (0-1 weeks after birth) was compared by using different age groups of bovine longissimus dorsi muscle tissue, different collagenase, different digestion time and different differential adhesion time. After 1.5 hours of collagenase type II digestion, more positive preadipocytes could be obtained by replacing with fresh medium 1.5 hours after seeding. At the same time, the use of immunofluorescence and other methods has further improved the reliability of the identification of intramuscular preadipocytes.

[0027]...

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Abstract

The invention relates to a method for separation, purification and identification of intramuscular adipocytes of Qinchuan beef cattle. Type II collagenase is used to digest the longissimus dorsi muscle of Qinchuan beef cattle, and the tissue block digested by type II collagenase is used to contain 10% fetal Neutralize the DMEM / F12 medium with bovine serum, filter and centrifuge to obtain the cells, wash and centrifuge the cells, resuspend them in DMEM / F12 medium containing 10% FBS, count them, and inoculate the cells in a culture dish , at CO 2 Incubate in the incubator for 1.5 hours, wash with sterile phosphate-buffered saline to remove non-adherent cells, and then culture the cells at 37°C, CO 2 In an incubator with a concentration of 5%, the medium was changed every 2 days, and after the confluence of the cells reached 70%-80%, positive cells were identified by immunofluorescence labeling. The intramuscular adipocytes obtained by the method have a higher positive ratio of pre-adipocytes, which lays a foundation for the study of increasing intramuscular fat deposition without increasing subcutaneous and visceral fat deposition.

Description

technical field [0001] The invention belongs to the technical fields of cell biology and cell engineering, in particular to a method for separating intramuscular fat cells of Qinchuan beef cattle. Background technique [0002] There are 4 major adipose tissue locations in livestock: visceral, subcutaneous, intramuscular and intermuscular. The accumulation of adipose tissue in the muscle is very important, and the excessive accumulation of adipose tissue in other locations is a burden on livestock production and greatly increases the cost of production, the common phenotype is that some animals are much larger than their counterparts, mainly not Accumulation of adipose tissue within the muscle, but primarily subcutaneous or visceral fat. However, to date, the mechanisms leading to the preferential accumulation of intramuscular fat remain poorly defined. [0003] Adipocytes, especially intramuscular adipocytes, share a common progenitor cell with myogenic cells during fetal ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/077G01N33/68
CPCG01N33/56966G01N33/68C12N5/0653C12N2509/00
Inventor 王晓宇昝林森李安宁张愈杨武才
Owner NORTHWEST A & F UNIV
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