Serum-free medium for porcine hepatocytes and preparation method of serum-free medium

Abstract

The invention relates to a serum-free medium for porcine hepatocytes and a preparation method of the serum-free medium. According to the serum-free medium for the porcine hepatocytes, a basic culture solution is added with the following components: 10.00mg / L of bovine insulin, 5.50mg / L of human transferrin, 5.02mg / L of sodium selenite, 2.00mg / L of ethanol amine, 196.26mug / L of dexamethasone, 0.5mg / L of fibronectin, 10.00mg / L of vitamin C, 60.00mug / L of hepatocyte growth factors, 80.00mug / L of epidermal growth factors, 2.00mg / L of glucagon and 1.50g / L of bovine serum albumin. The serum-free medium for the porcine hepatocytes is capable of being used for culturing porcine primary hepatocytes, the porcine primary hepatocytes are very equivalent to that cultured by a serum-containing medium in form and vigor and the growth condition is obviously good; the original biological characteristics and biological functions of the porcine hepatocytes can be well kept; the negative effects caused by a traditional serum-containing medium can be avoided; the condition requirements of normal growth and division and proliferation of the hepatocytes can be met.

Description

technical field [0001] The invention relates to the technical field of animal cell culture, in particular to a serum-free culture medium for pig hepatocytes and a preparation method thereof. Background technique [0002] The liver is an important organ for human and animal bodies to store glycogen, regulate metabolism, deoxidize, secrete active factors, and biotransform, and it is also the main site for drug metabolism and detoxification. Liver cells are the main executors of the above-mentioned biological functions of liver tissue. For this reason, many in vitro experiments often use primary human or livestock hepatocytes as research models to develop the physiological regulation function of liver cells, metabolic diseases, and pharmacology. , Molecular mechanisms of pathology and toxicology, establishment of virus-infected cell models, and research in the fields of physical and chemical factors and toxic effects. The in vitro culture of primary hepatocytes is the first st...

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Problems solved by technology

In addition, some researchers have used other serum-free media for related research, but these serum-free media contain animal-derived components and cannot be used for suspension culture of porcine primary liver cells, which is difficult to meet the needs of scientific research and clinical applications

Other commercial hepatocyte serum-free media have many problems, such as the secret formula of the media, high price, and uncertain preparation methods, which make it difficult to popularize and apply in the field of serum-free culture of hepatocytes in vitro

Although some domestic patent applications disclose some serum-free hepatocyte culture medium, the developed serum-free hepatocyte culture medium can be applicable to the bioartificial liver support system, but it is difficult to be used or suitable for the cultivation of porcine hepatocyte serum-free. The research and application of the latter cannot be satisfied, so it is urgent to develop a new serum-free medium for porcine hepatocytes

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