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Ear margin tissue fibroblast line of Minxian county black fur sheep and its culture process

A fibroblast and culture method technology, applied in tissue culture, animal cells, vertebrate cells, etc., can solve the problems of reducing the number of varieties, reducing the number of varieties, improper preservation, etc., and improving the cell purity and the quality of cryopreserved cells. Stable, cost-reducing effect

Inactive Publication Date: 2007-08-22
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

In most developed countries, with the intensification of animal husbandry production systems, only a few breeds and hybrids with high economic value are raised in large quantities, and the number of breeds is rapidly decreasing; in some developing countries, although there are relatively rich genetic resources, but The number of original local varieties has been greatly reduced due to improper conservation of species and blind introduction of foreign varieties for hybridization

Method used

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  • Ear margin tissue fibroblast line of Minxian county black fur sheep and its culture process
  • Ear margin tissue fibroblast line of Minxian county black fur sheep and its culture process
  • Ear margin tissue fibroblast line of Minxian county black fur sheep and its culture process

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0127] Example 1 Construction of high cell viability and high purity Minxian black fur sheep ear margin tissue fibroblast cell line

[0128] (1) Primary culture: Under aseptic conditions, scrape off the hair on the surface of the ear margin tissue samples of Minxian black fur sheep with ophthalmic scissors, put them into a petri dish filled with PBS and rinse them for 6-8 times to remove surface blood stains and Hair; cut tissue into 1 mm 3 Left and right tissue pieces; use a pipette to transfer the tissue pieces into the culture flasks to disperse evenly, invert the culture flasks, and incubate at 37°C, 5% CO 2 CO 2 Cultivate in the incubator for 2-3 hours; add 8ml-10ml of medium after the tissue block is completely attached, medium composition: DMEM+8% special grade fetal bovine serum, continue to cultivate for 10-12h;

[0129] (2) Subculture: Discard the culture medium in the culture flask in step (1), wash the residue in the culture flask twice with PBS to remove residua...

Embodiment 2

[0139] Example 2 Detection of Biological Characteristics of Minxian Black Fur Sheep Ear Tissue Fibroblast Cell Line

[0140] The biological characteristics of the Minxian black fur sheep ear margin tissue fibroblast cell line cultured in Example 1 were examined, and the methods and conclusions are as follows. 1. Observation of cell morphology

[0141] 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.

[0142] Conclusion: Figure 1 (primary cells of Minxian black fur sheep), Figure 2 (cells of Minxian black fur sheep before subculture), Figure 3 (cells of Minxian black fur sheep subculture 1), Figure 4 (cells of Minxian black fur sheep As shown in sheep embryo subculture II cells), when the cells grow well, their morphology is typical fibroblast, presenting fusiform or irregular triangle. Observing the overall picture of the ...

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Abstract

The present invention is high activity and high purity fibroblast line of Minxian county black fur sheep obtained with the ear margin tissue and through primary cell culture, subculture and cell freezing preservation and in the preservation number of CGMCC No.1880, and belongs to the field of cell biology. The high activity and high purity fibroblast line is suitable for great scale culture. The present invention makes it possible to provide great amount of high quality fibroblast line of Minxian county black fur sheep for the scientific research in gene engineering, cell engineering, etc.

Description

technical field [0001] This invention is about the establishment of a cell line and its culture method, that is, the ear margin tissue of Minxian black fur sheep is used for cell separation, primary culture, and subculture to obtain high survival rate and high purity Minxian black fur Sheep ear margin tissue fibroblast cell line. The present invention belongs to the field of biology. Background technique [0002] Genetic resources of livestock and poultry are an important part of biodiversity, and also the part that has the closest and most direct relationship with human beings. Globally, genetic resources of livestock and poultry provide more than 30% of food and agricultural products for human beings. According to the predictions of the governments of developed countries and the World Food and Agriculture Organization, 90% of the varieties of global agriculture will be provided through molecular breeding in this century, and the contribution rate of varieties to the entir...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06C12N5/071
Inventor 马月辉关伟军张艳艳马忠仁陈莉娜刘长青
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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