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Method for separating 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: 2019-03-19
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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  • Method for separating intramuscular fat cells of Qinchuan beef cattle
  • Method for separating intramuscular fat cells of Qinchuan beef cattle
  • Method for separating intramuscular fat cells of Qinchuan beef cattle

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Embodiment Construction

[0025] During the research process, the applicant found through experimental comparison that using type II collagenase to digest the longissimus dorsi muscle of Qinchuan beef calves can obtain more positive preadipocytes, compared with other methods such as type I collagenase digestion, or with adult cattle There are some differences in the intramuscular fat cells obtained from the longissimus dorsi muscle.

[0026] The applicant compared the longissimus dorsi muscle tissue of newborn cattle (0-1 week old) by using different collagenases, different digestion times, and different differential attachment times. After 1.5 hours of collagenase type II digestion, more positive preadipocytes can be obtained by replacing the fresh medium 1.5 hours after inoculation. At the same time, the reliability of identifying intramuscular preadipocytes was further improved by using methods such as immunofluorescence.

[0027] In the following examples, the involved animal tissues and reagent s...

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Abstract

The invention relates to a method for separating, purifying and identifying intramuscular fat cells of Qinchuan beef cattle. The method comprises the following steps: digesting longissimus dorsi of the Qinchuan beef cattle with type II collagenase, neutralizing tissue blocks digested by the type II collagenase by using a DMEM / F12 medium containing 10% of fetal bovine serum, filtering and centrifuging to obtain cells; washing and centrifuging the cells, re-suspending the cells in the DMEM / F12 medium containing 10% of FBS (Fetal Bovine Serum), counting, inoculating the cells in a culture dish, culturing in a CO2 incubator for 1.5 hours, and washing to remove non-adherent cells with a sterile phosphate buffer solution; culturing the cells in an incubator having a CO2 concentration of 5% at 37DEG C, changing the medium once every 2 days, and performing cellular immunofluorescence labeled positive cell identification after the cell confluence degree reaches 70-80%. The intramuscular fat cells of the beef cattle obtained by the method in the invention have high a preadipocyte positive rate, and a foundation is laid for study of not increasing subcutaneous and visceral fat deposition while improving intramuscular fat deposition.

Description

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

Claims

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Application Information

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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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