Shanghai Jinma analyzes for you: the problem of adding animal serum during animal cell culture
In recent years, especially this year's Zhejiang college entrance examination test questions related to the content of cell culture, it is necessary to appropriately expand the knowledge of cell culture.
The textbook says that animal cell clone culture should be added with animal serum (fetal bovine serum is the best). The reason is that in addition to providing the necessary ingredients, it is also related to the internal environment of cell life. Serum can create the internal environment of cell life, such as certain Stability of osmotic pressure and pH. So is it necessary to add animal serum? According to the data, adding serum has its importance:
The synthetic medium can only maintain the survival of the cells. To grow and reproduce the cells, a certain amount of natural medium (such as serum) needs to be added.
The serum contains:
â‘ Various proteins (albumin, globulin, ferritin, etc.)
â‘¡Various metal ions;
â‘¢ hormone;
â‘£Promotes the attachment of substances such as fibronectin, cryoglobulin, collagen, etc.
⑤Various growth factors
â‘¥transfer protein
⑦Unknown ingredients
Generally speaking, the medium containing 5% calf serum can keep the cells from dying for most cells, but to support the growth of cells, it is necessary to add 10% serum.
The biological effects on serum supporting cell growth have been proven, but the complex components in serum have not been fully understood so far.
Not only does the cell growth factor exist in the serum, but there are also cell growth inhibitory factors or toxic factors in the same pair. Therefore, the biological characteristics reflected by the cells cultured in the serum-containing medium are the comprehensive reaction of cells and complex serum factors.
The quality of the serum is the key to the success of the experiment. Commonly used serums include fetal bovine blood cleanse, newborn bovine serum, calf blood, rabbit serum, horse serum, etc., of which fetal bovine serum is the best quality. The standard of high-quality serum: transparent, light yellow, no sediment, no bacteria, mycoplasma, virus contamination. Serum inactivation (elimination of complement activity): 56 ° C for 30 minutes. Serum disinfection: filter sterilization
However, in the research fields of growth factors, protein engineering, gene expression regulation, etc., there is an urgent need to cultivate cells with serum-free medium.
The main development strategy of serum-free medium: supplement the basic medium with various essential factors, such as hormones, growth factors, binding proteins, adherence and expansion factors. Serum-free medium consists of basic medium and supplementary components to replace serum.
In 1975, Sato successfully cultivated pituitary cell lines with serum-free medium for the first time. In the past 20 years, dozens of cell lines have been reported to successfully grow and proliferate in serum-free medium.
The use of serum-free cell culture medium ensures the accuracy, repeatability and stability of the experimental results, reduces cell contamination, and simplifies the procedures for purification and identification of various cell products.
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