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Mount Wuzhi Pig ear rim tissue fibroblast series and culturing method thereof

A technology of fibroblasts and culture methods, applied in tissue culture, animal cells, vertebrate cells, etc., can solve the problems of low cell activity and purity, high cell death rate, single method, etc., and achieve stable cell quality and growth , cost reduction effect

Inactive Publication Date: 2007-08-08
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For the preservation of animal cells, there are high requirements for cell purity and activity. At present, the adherent culture method also has some shortcomings, such as single method, high cost, low cell activity and purity, and low cell activity after cryopreservation and recovery. high mortality rate

Method used

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  • Mount Wuzhi Pig ear rim tissue fibroblast series and culturing method thereof
  • Mount Wuzhi Pig ear rim tissue fibroblast series and culturing method thereof
  • Mount Wuzhi Pig ear rim tissue fibroblast series and culturing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0137] Example 1 Construction of High Viability and High Purity Wuzhishan Minipig Ear Marginal Tissue Fibroblast Cell Line

[0138] (1) Primary culture: Wash the harvested Wuzhishan minipig ear margin tissue with PBS plus 1% double antibody for 6-8 times and cut it into 0.5-1.5 mm 3 The tissue block; move the tissue block into the culture bottle and spread it on the bottom of the bottle, and place it upside down at 37°C, 5% CO 2 Culture in an incubator for 3 to 4 hours; add 8 to 10 ml of culture medium after the tissue block is completely adhered to the wall, and the composition of the culture medium is: DMEM+10% special grade fetal bovine serum.

[0139] (2) Subculture: Discard the culture medium in the culture flask in step (1), and carefully wash the residue in the culture flask twice with PBS preheated to 37°C to remove residual serum, exfoliated tissue pieces and dead cells . After digesting with trypsin, add 6-10ml of culture medium to stop the digestion; the digested ...

Embodiment 2 5

[0149] Example 2 Detection of biological characteristics of Wuzhishan minipig ear margin tissue fibroblast cell line

[0150] The biological characteristics of the Wuzhishan minipig ear margin tissue fibroblast cell line cultured in Example 1 were examined, and the methods and conclusions are as follows.

[0151] 1. Observation of cell morphology

[0152] Methods: In the process of cell culture in vitro, the cells were routinely inspected to observe the growth status of the cells, the color change of the culture medium and whether the cells were polluted.

[0153] Conclusion: Figure 1 (primary cells of Wuzhishan minipigs), Figure 2 (cells of Wuzhishan minipigs before subculture), Figure 3 (cells of Wuzhishan minipigs subculture 1), and Figure 4 (cells of Wuzhishan minipig embryos subcultured II) , when the cells grow well, their morphology is typical fibroblast, presenting fusiform or irregular triangle. Observing the overall picture of the cell population, the cultured cell...

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Abstract

The invention discloses a high-living rate and high-purity mature fiber cell system of Mongolian horse (CGMCC No.1879) in the cellar biological domain, which is characterized by the following: culturing high-purity phoroblast without epithelial cell; maintaining the living rate of frozen cell between 93.2% and 98.7%; fitting for large scale of culturing.

Description

technical field [0001] The present invention relates to a cell line and its culture method. Specifically, it uses Wuzhishan minipig ear margin tissue for primary culture and subculture to obtain a high-survival, high-purity Wuzhishan miniature pig ear margin tissue fibroblast cell line, which belongs to cell biology. field of study. Background technique [0002] The genetic resources of livestock and poultry are important biological resources for the survival and development of human society, and are important strategic basic resources of the country. They are related to national security and sovereignty, and have become one of the focuses of competition in the world "bio-economy" era. [0003] In recent years, due to the continuous deterioration of the biological living environment, biological germplasm resources are facing a serious crisis, which directly threatens the survival and development of human beings. According to the 1995 statistics of the Food and Agriculture O...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06C12N5/071
Inventor 关伟军马月辉刘学东刘帅白修云李向臣
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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