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Method for preparing culture medium without blood serum for epidermis stem cell of milch goat in the central Shanxi plain

A serum-free medium, epidermal stem cell technology, used in biochemical equipment and methods, tissue culture, animal cells, etc.

Inactive Publication Date: 2005-12-21
NORTHWEST A & F UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, there is no serum-free medium suitable for long-term expansion and culture of purified epidermal stem cells at home and abroad.

Method used

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  • Method for preparing culture medium without blood serum for epidermis stem cell of milch goat in the central Shanxi plain
  • Method for preparing culture medium without blood serum for epidermis stem cell of milch goat in the central Shanxi plain
  • Method for preparing culture medium without blood serum for epidermis stem cell of milch goat in the central Shanxi plain

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Embodiment Construction

[0017] The present invention will be described in further detail below in conjunction with the examples given by the inventor.

[0018] Referring to the accompanying drawings, the key of the present invention is the production of fibroblast conditioned medium and the screening of reasonable concentrations of various factors. The production of serum-free medium for epidermal stem cells of Guanzhong dairy goat is now taken as an example to illustrate.

[0019] 1. Preparation of fibroblast conditioned medium: Aseptic Guanzhong dairy goat neck skin was treated with 0.25% Trypsin+0.02% EDTA (overnight at 4°C) to separate the epidermis from the dermis. Wash the cells between the dermis and epidermis, cut the dermis into small pieces and paste them in glass dishes (Φ=50mm). 3-4 pieces per plate were added with culture medium (F12+1%BSA+10ng / ml EGF+10ug / ml insulin+10ng / mlIGF-1+green streptavidin 100u / ml) for cultivation. When the fibroblasts covered the plate, the tissue pieces were...

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Abstract

The invention discloses a preparation method of a serum-free medium for epidermal stem cells of Guanzhong dairy goats, including the screening of fibroblast conditioned medium and the appropriate concentration of IGF-1, EGF, and GSFCM, and finally determines the formula of the medium: F12+1-1.2 %BSA+20~25%GSFCM+20~25ng / mlEGF+5ug / ml insulin+15~20ng / ml IGF-1+0.4~0.6μg / ml hydrocortisone+penicillin 100u / ml; the present invention Breaking through the limitation of adding only epidermal growth factor to the traditional medium, adding insulin-like growth factor, and finding out the appropriate ratio of this factor, adopting the method of adding fibroblast conditioned medium to the medium instead of co-culture with fibroblasts Methods. The present invention uses hormone, cytokine combination, fibroblast conditioned medium and BSA to supplement the lack of nutritional factors caused by not adding serum, and fully meets the nutritional needs of epidermal stem cells for long-term expansion and culture in vitro. Commercially produced serum-free medium.

Description

technical field [0001] The invention relates to the field of in vitro culture medium for stem cells, in particular to a method for preparing a serum-free culture medium for Guanzhong dairy goat epidermal stem cells. Background technique [0002] Cell culture is an important link in the production of genetically engineered products. Because the traditional serum-containing cell culture will bring many unfavorable factors to production and scientific research, since the 1950s, foreign countries have begun to develop serum-free medium. In the late 1980s and early 1990s, serum-free media with commercial application value were launched one after another. One of the main characteristics of serum-free media is that the applicable cell lineage is narrow. For specific cell lines, it is necessary to find out new formulas or optimal formulas for specific cell lines. condition. Epidermal stem cells are the source of cells to maintain the structural and fu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071
Inventor 杨学义窦忠英
Owner NORTHWEST A & F UNIV
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