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Method for performing in-vitro culture to bovine preadipocytes

A fat cell and in vitro culture technology, applied in the field of cell engineering and tissue engineering, can solve the problems of manpower consumption, achieve high efficiency, scientific and reasonable medium preparation, and shorten the processing time

Inactive Publication Date: 2012-07-11
INST OF ANIMAL SCI & VETERINARY MEDICINE SHANDONG ACADEMY OF AGRI SCI
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Problems solved by technology

After 1 day of culture, a small number of cells begin to adhere to the wall, and after 7-8 days, about 80-90% of the cells adhere to the wall. Therefore, this method saves time compared with the tissue block culture method, but it is also labor-intensive when cutting the tissue

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  • Method for performing in-vitro culture to bovine preadipocytes
  • Method for performing in-vitro culture to bovine preadipocytes
  • Method for performing in-vitro culture to bovine preadipocytes

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Embodiment 1

[0020] (1) Reagent

[0021] 0.1% Collagenase I medium: Dissolve 100 mg collagenase I in 100 ml D-Hanks solution (model: 91105B-100, produced by Shanghai Baoman Biotechnology Co., Ltd.), and use 5.6% NaHCO 3 The pH value of the solution was adjusted to 7.2-7.4, filtered through a 0.22 micron filter, subpackaged, and stored at -20°C for later use.

[0022] Cell culture medium: add 10% fetal calf serum by volume to DMEM / F12 solution (model: D6421, produced by Sigma Company), filter through a 0.22 micron filter, aliquot, and store at 4°C for later use.

[0023] (2) Primary culture of bovine preadipocytes

[0024] a. Under aseptic conditions, take the subcutaneous fat tissue from the bovine, transfer it to a plate filled with D-Hanks solution, rinse it 6 times, and use ophthalmic scissors to remove the visible blood vessels and connective tissue in the fat tissue;

[0025] b Then cut the tissue block to 9mm 3 The small pieces were homogenized to a chylus shape with a tissue homo...

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Abstract

The invention discloses a method for performing in-vitro culture to bovine preadipocytes, belonging to the technical field of cell and tissue engineering. The method is based on the traditional subculture method, a tissue homogenizer is utilized and combines with the collagenase I culture medium to treat the fatty tissue, a DMEM (Dulbecco's Modified Eagle Medium) / F12 culture solution containing fetal bovine serum is used to perform in-vitro culture to bovine preadipocytes, and a new model of the in-vitro culture of bovine preadipocytes is successfully built. The technology is advanced and simple, the preparation of the culture medium is scientific and reasonable, manpower and material resources are saved, and the treatment time of the test sample is greatly shortened; the obtained cell morphology is best, the RNA (Ribonucleic Acid) culture and extraction efficiency is maximum, the preadipocyte culture method is convenient and efficient in a long term and lays the foundation for the gene expression and protein expression and the signal pathway in the later intensive study of the proliferation and differentiation process of preadipocytes; and a new idea and a new method are provided to regulate body fatness deposition of beef cow and prevent diseases in the perinatal period and lactation peak period of milk cow, etc.

Description

technical field [0001] The invention relates to a method for culturing bovine preadipocytes in vitro, belonging to the fields of cell engineering and tissue engineering. Background technique [0002] Cells isolated and cultured from adipose tissue are often called "preadipocytes". Preadipocytes are a type of adipocyte precursors with the ability to proliferate and differentiate. Under the influence of various factors in the body, they differentiate into mature adipocytes. Currently, the successfully cultured preadipocytes are mainly derived from human, mouse, pig and so on. In 1995, Aso et al isolated and cloned a preadipocyte cell line from SV (stromal vascular) cell culture between bovine muscle cells. In recent years, some domestic laboratories have also carried out research on bovine fat cells. The currently used culture methods mainly include tissue block culture method and traditional digestion culture method. When using the tissue block culture method, the adipose...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077
Inventor 刘晓牧宋恩亮刘桂芬成海建谭秀文万发春游伟
Owner INST OF ANIMAL SCI & VETERINARY MEDICINE SHANDONG ACADEMY OF AGRI SCI
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