Primary isolated culture and induced differentiation method for precursor fat cells in adult yak muscles

A technique for separating and culturing adipocytes, which is applied in the field of cell engineering and tissue engineering, can solve the problems of high pollution rate and low activity, and achieve the effects of avoiding pollution, reducing cell damage, and shortening the test time

Active Publication Date: 2021-11-30
SICHUAN AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003]Currently, there are no reports and studies on the in vitro culture technology of yak intramuscular preadipocytes, and the isolation techniques of other adult animal intramuscular preadipocytes generally have low activity and high The problem of pollution rate, in order to solve the former problem, most of the sampling objects are young animals, but high pollution is still a major problem in primary cell culture experiments, and there is no effective solution

Method used

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  • Primary isolated culture and induced differentiation method for precursor fat cells in adult yak muscles
  • Primary isolated culture and induced differentiation method for precursor fat cells in adult yak muscles
  • Primary isolated culture and induced differentiation method for precursor fat cells in adult yak muscles

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Embodiment 1

[0030] Embodiment 1: A method for primary isolation, culture and differentiation of adult yak intramuscular preadipocytes, comprising the following steps:

[0031] S1. Disinfection treatment: Isolate the longissimus dorsi muscle tissue of adult yaks under aseptic conditions, rinse the surface with PBS until there is no dirt, soak in iodine for 10 minutes, and then quickly remove and peel off the outer stained area tissue;

[0032] S2. Segmentation: Rinse the sterilized tissue with PBS and divide it into small pieces, put it into a 50mL centrifuge tube containing 2mL 5% PBS and cut it into 1-2mm in size 3 organization block;

[0033] S3. Digestion: Add a digestive enzyme solution to the centrifuge tube containing the tissue block, the volume ratio of the digestive enzyme solution to the tissue block is 3:1, and digest at a constant temperature of 37°C for 120 minutes to obtain a digestive juice; wherein, the digestive enzyme solution is 100mg type I collagenase, 50mg type II c...

Embodiment 2

[0038] Embodiment 2: A method for primary isolation, culture and differentiation of adult yak intramuscular preadipocytes, comprising the following steps:

[0039] S1. Disinfection treatment: Isolate the longissimus dorsi muscle tissue of adult yaks under aseptic conditions, wash the surface with PBS until there is no dirt, soak in iodine for 15 minutes, and then quickly remove and peel off the outer stained area tissue;

[0040] S2. Segmentation: Rinse the sterilized tissue with PBS and divide it into small pieces, put it into a 50mL centrifuge tube containing 2mL 5% PBS and cut it into 1-2mm in size 3 organization block;

[0041] S3. Digestion: Add a digestive enzyme solution to the centrifuge tube containing the tissue block, the volume ratio of the digestive enzyme solution to the tissue block is 4:1, and digest at a constant temperature of 37°C for 150 minutes to obtain a digestive juice; wherein, the digestive enzyme solution is 100mg type I collagenase, 50mg type II co...

Embodiment 3

[0046] Embodiment 3: A method for primary isolation, culture and differentiation of adult yak intramuscular preadipocytes, comprising the following steps:

[0047] S1. Disinfection treatment: separate adult yak longissimus dorsi muscle tissue under aseptic conditions, wash the surface with PBS until there is no dirt, soak in iodine for 8 minutes, and then quickly remove and peel off the outer stained area tissue;

[0048] S2. Segmentation: Rinse the sterilized tissue with PBS and divide it into small pieces, put it into a 50mL centrifuge tube containing 2mL 5% PBS and cut it into 1-2mm in size 3 organization block;

[0049] S3. Digestion: Add a digestive enzyme solution to the centrifuge tube containing the tissue block, the volume ratio of the digestive enzyme solution to the tissue block is 2.5:1, digest at a constant temperature of 37°C for 100 minutes, and obtain a digestive juice; wherein, the digestive enzyme solution is 100mg type I collagenase, 50mg type II collagenas...

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Abstract

The invention discloses a primary isolated culture and induced differentiation method for precursor fat cells in adult yak muscles. The method comprises the steps of disinfection treatment and segmentation, digestion, screening and filtration, centrifugation, purification and culture. According to the cell culture for the precursor fat cells in the adult yak muscles, provided by the invention, manpower and material resources can be saved, and the limitation of high pollution rate caused by using newborn calf fat tissues and primary cells in a traditional culture method is broken through, so that the source approach of samples is wider, and a large number of cells can be obtained at the same time; and the culture method is a convenient and efficient precursor fat cell culture method, provides a molecular basis for subsequent in-depth research on gene and protein expression, signal channels and the like in proliferation and differentiation process of intramuscular precursor fat cells, provides a new thought and a new method for research on fat deposition, fat metabolism, fat mobilization and the like of different parts, and has an important significance in improving the meat quality of yaks.

Description

technical field [0001] The invention belongs to the field of cell engineering and tissue engineering, and in particular relates to a method for primary isolation, culture and differentiation induction of intramuscular preadipocytes of adult yaks. Background technique [0002] Yak is a unique breed of cattle in China, which can adapt to the harsh natural environment in plateau areas. Moreover, yak has high economic value and is an important means of life and production for local herdsmen to survive. Adipose tissue is mainly distributed in four parts: internal organs, subcutaneous, intramuscular, and intermuscular. Exploring the accumulation rules of adipose tissue in each part is of great significance for improving the quality of yak meat and for further increasing the economic income of local herdsmen. However, the way of raising yaks is backward, and most of them adopt the traditional grazing production method. Although studies in recent years have proved that yaks grow we...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077
CPCC12N5/0653C12N2509/00C12N2509/10C12N2500/34
Inventor 王之盛胡瑞王森师俊华邹华围王俊梅郭逸芯
Owner SICHUAN AGRI UNIV
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