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Tianzhushan white yak ear-edge tissue fiber cell system and constituting method

A fibroblast and yak technology, applied in the field of cell biology, can solve the problems of cell viability and purity not meeting the required standards, complicated operation steps, unsuitability, etc. Effect

Inactive Publication Date: 2007-08-29
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

[0011] At present, Epstein-Barr virus transformation method, adherent culture method and enzyme digestion method are often used for mass culture of poultry species in vitro cells, but the results of comparative experiments show that: EB virus transformation method is not suitable for in vitro cell culture of poultry species, and its cultured The cell viability and purity cannot meet the required standards. The effect of the EB virus transformation method on human and primate cell culture is better than that of poultry cell culture; the primary cells obtained by enzyme digestion method and adherent culture method are passaged After inoculation, the cell population doubling time (PDT) was 35.9h and 48h respectively
However, the former has complicated operation steps and is not suitable for large-scale cell culture in vitro, while the latter is simple and easy, and is suitable for large-scale cell culture in vitro

Method used

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  • Tianzhushan white yak ear-edge tissue fiber cell system and constituting method
  • Tianzhushan white yak ear-edge tissue fiber cell system and constituting method
  • Tianzhushan white yak ear-edge tissue fiber cell system and constituting method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0158] Example 1 Construction of high cell viability and high purity Tianzhushan white yak ear tissue fibroblast cell line

[0159] (1) Primary culture:

[0160] Under sterile conditions, place the ear margin tissue block of white yak collected from Tianzhu Mountain in Tianzhu County in a small petri dish, scrape off the uncut hair on the surface of the tissue block, rinse it with PBS for 3 to 4 times, and then cut it. into 0.5~1.5mm 3 the tissue block; transfer the tissue block into the culture flask, invert the culture flask, and store in saturated humidity, 37°C, 5% CO 2 Incubate in a carbon dioxide incubator for 2 to 3 hours; add 6 to 10 ml of culture medium (the culture medium is DMEM supplemented with 10% fetal bovine serum, 0.5 μmol / L sodium selenite) after the tissue pieces are completely adhered to the wall;

[0161] (2) Subculture: discard the culture medium in the culture flask in step (1), wash the residues in the culture flask with PBS twice, digest with trypsin...

Embodiment 2

[0171] Example 2 Detection of biological characteristics of fibroblast cell line in Tianzhushan white yak ear tissue

[0172] The biological characteristics of the Tianzhushan white yak ear margin tissue fibroblast cell line cultured in Example 1 were examined, and the methods and conclusions were as follows.

[0173] 1. Cell morphology observation

[0174] Methods: During the cell culture in vitro, the cells were routinely checked to observe the cell growth state, the color change of the culture medium and whether the cells were contaminated.

[0175] Conclusion: As shown in Figure 1 and Figure 2, when the cells grow well, their morphology is typical fibroblastic, fusiform or irregular triangle. The overall observation of the cell population shows that the cultured cells show a radial, flame or swirling trend, which proves that the cells grow healthily and vigorously.

[0176] 1. Bacteria and fungi detection

[0177] method:

[0178] (1) Take the cryopreserved cells with ...

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Abstract

The invention relates to a method for establishing and culturing Tianzhu Yak'ear marginal fibroblast cell line, utilizing the fast tissue wall pasting method to improve culture medium, and finally obtaining high-yield and high-purity Tianzhu Yak'ear marginal fibroblast cell line whose conservation number is CGMCC No.1881, belonging to the field of cell biology. And the frozen cells have stable quality, suitable to large scale culture. And the Tianzhu Yak'ear marginal fibroblast cell line can provide large numbers of high quality materials for research on science of living systems in gene engineering, cell engineering, immunity and molecular biology; can act as donor cells for animal's somatic cell cloning and breeding; and can enrich local variety improvement and storage of local excellent variety in agriculture.

Description

technical field [0001] The invention relates to a Tianzhushan white yak ear margin tissue fibroblast cell line and a construction method thereof, in particular to the use of Tianzhushan white yak ear margin tissue cells for primary culture and subculture to obtain high viability and high purity Tianzhushan white yak ear margin tissue A fibroblast cell line belongs to the field of cell biology. Background technique [0002] In order to ensure the sustainable development of human society, the United Nations held the "World Conference on Environment and Development" in Rio de Janeiro, the capital of Brazil, in June 1992, at which heads of countries signed the "Convention on Biological Diversity". As of February 2004, a total of 188 countries around the world have signed the Convention, and my country is one of the first signatories. This shows that the vast majority of countries in the world have recognized that the issue of biodiversity is a major issue for all mankind. Sinc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06C12N5/071
Inventor 马月辉关伟军李向臣娜日苏于太永刘鹏
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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