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Cell culture means that cells grow in vitro, and animal cells no longer form individuals in the process of individual cell culture.

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concept

Animalcellculture is to take out the relevant tissues in animals, disperse them into single cells (using trypsin or collagenase), and then put them into a suitable culture medium to allow these cells to grow and proliferate.

brief introduction

1. Composition of animal cell culture solution: The nutrients required for cell culture in vitro are basically the same as those in vivo, such as sugar, amino acids, inorganic salts, growth promoting factors and trace elements. A culture medium which is formed by strictly mixing the above substances required by cells according to their types and required amounts is called a synthetic culture medium. Because there are still some components in the internal environment where animal cells live, it is necessary to add animal serum to provide an environment similar to that in vivo. Therefore, when using synthetic culture medium, it is usually necessary to add some natural ingredients such as serum and plasma. [ 1]

2. Characteristics of animal cell culture medium: liquid culture medium, usually containing animal serum.

3. Conditions for animal cell culture:

① Sterile and nontoxic environment: sterilize the culture solution and all the culture vessels, and usually add a certain amount of antibiotics to the culture solution to prevent pollution. In addition, the culture medium should be changed regularly to remove metabolites and prevent the accumulation of cell metabolites from harming the cells themselves.

② Nutrients: inorganic substances (inorganic salts, trace elements, etc. ) and organic substances (sugar, amino acids, growth promoting factors, etc. ).

Serum and plasma, but serum and plasma are not nutrients.

③ Temperature and pH (36.5 0.5℃, 7.2 ~ 7.4)

④ Gas environment (mixed gas of 95% air and 5% CO 2)

Among them, 5%CO2 gas is used to keep the pH value of the culture solution stable.

3. Basic process

Take organs and tissues of animal embryos or young animals. The material is cut into pieces, treated (digested) with trypsin (or collagenase) to form dispersed single cells, and the treated cells are transferred to a culture medium to make a cell suspension with a certain concentration. The dispersed cells in the suspension will quickly attach to the bottle wall and become cell adhesions. When adherent cells divide and grow into contact with each other, cells will stop dividing and proliferating, and contact inhibition will occur. At this time, the contact-inhibited cells need to be treated with trypsin again. Then prepare a cell suspension with a certain concentration. In addition, primary culture is the cell and tissue culture immediately after being taken out of the body. When cells grow and migrate from animals and plants, forming growth halos and increasing, scientists carry out subculture, that is, divide the primary cultured cells into several parts and inoculate them into several media to make them continue to grow and proliferate. Cells with special properties obtained from primary cultures or cell lines through certain selection or purification methods are called cell lines. After more than 50 generations of culture, most cells have aged and died, but some cells have changed their genetic material, which has the characteristics of unlimited passage, that is, canceration. Cells at this time are called cell lines.