Milk cow mammary glandular cell in vitro isolated culture method

A technique for separating and culturing mammary epithelial cells, which is applied in the field of cell separation and culture in vitro, and can solve the problems of different trypsin sensitivities.

Inactive Publication Date: 2008-11-12
NORTHEAST INST OF GEOGRAPHY & AGRIECOLOGY C A S
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Problems solved by technology

Its shortcoming is that mammary gland cells of different animal species have different sensitivities to trypsin, so determining the o

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Embodiment Construction

[0009] The present invention will be further described below in conjunction with the actual operation steps of the specific implementation.

[0010] (1) Primary culture of dairy cow mammary gland tissue

[0011] The mammary gland tissues of healthy black-and-white dairy cows slaughtered locally on the same day were taken and brought back to the laboratory in an ice box. The mammary gland tissue was sterilized by soaking in 75% alcohol. In the ultra-clean workbench, the mammary gland tissue block was transferred to a plate with cell culture medium, cut into small pieces with a diameter of about 3mm, and the culture medium was sucked up with a straw for repeated washing. Use tweezers to separate connective tissue and adipose tissue as much as possible, then cut into small pieces with a diameter of about 1 mm, and insert them into a petri dish with a distance of about 0.5 cm between each small piece. To prevent the tissue pieces from drying out, add 2 drops of cell growth soluti...

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Abstract

The invention discloses a method for isolating and cultivating milk cow breast cells in vitro, relating to the cell in vitro isolation and culture technology field and solving the problem that breast fibrocytes and epithelial cells of a milk cow cannot be isolated completely during the isolation and culture of the breast epithelial cells of the milk cow. The method comprises the following steps of: selecting mammary tissue of a milk cow which is killed on the very day from a local abattoir, adopting tissue explants technique and graded trypsin digestion technique to isolate and cultivate the breast epithelial cells of the milk cow according to the different sensibility of the fibroblasts and epithelial cells to trypsin and the different adhering time, purifying the breast epithelial cells of the milk cow, identifying with keratin 18 to obtain the corresponding cell system finally. The method provides the best trypsin concentration when isolating and cultivating the breast epithelial cells of a milk cow, and can completely isolate the breast fibrocytes and the epithelial cells of the milk cow from each other.

Description

technical field [0001] The invention relates to a method for separating and culturing cells in vitro, in particular to a method for separating and culturing dairy cow mammary gland cells in vitro. Background technique [0002] The mammary gland is a multiple-tube vesicular skin gland. The mammary gland epithelium has a special milk secretion function. Many components of milk can only be synthesized by mammary gland epithelial cells. Therefore, the isolation and culture of mammary gland epithelial cells is one of the important means to study mammary gland function and its regulation. . Breast cell culture has a history of half a century. Since Lasfargues (1957) used collagenase digestion to separate breast cells from breast tissue, many scientists have carried out extensive research. [0003] In vitro cell separation and culture technology is a basic method to reproduce the main aspects of cell life activities in vitro. Since its establishment, it has been applied to almost ...

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Application Information

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IPC IPC(8): C12N5/06C12N5/071
Inventor 刘春龙李忠秋孙海霞李长胜
Owner NORTHEAST INST OF GEOGRAPHY & AGRIECOLOGY C A S
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