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Culture medium for culturing hepatocytes and preparation method thereof

A medium and hepatocyte technology, applied in biochemical equipment and methods, tissue culture, animal cells, etc., can solve the problems of unfavorable daily application of hepatocyte industrial culture and high price, and achieve the purpose of promoting cell adhesion, proliferation, With a reasonable effect

Inactive Publication Date: 2017-02-22
叶宗耀
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, although a serum-free medium for hepatocyte culture has been developed, it is expensive, which is not conducive to daily application, widespread promotion, and industrialized culture of hepatocytes.

Method used

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  • Culture medium for culturing hepatocytes and preparation method thereof
  • Culture medium for culturing hepatocytes and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1, a kind of medium for cultivating hepatocytes

[0028] The medium for cultivating hepatocytes includes basal medium and additives added in the basal medium, and the ingredients of the additives include: cholesterol 4mg / L, lipoic acid 0.2mg / L, cell Growth factor 20μg / L, chondroitin sulfate 0.2mg / L, biotin 13μM, insulin 0.9mg / L, catalase 10mg / L, taurine 0.1g / L, myristic acid 6mg / L, fibronectin 14μg / L.

[0029] The basal medium is F12 medium.

[0030] The cell growth factor is composed of epidermal growth factor and hepatocyte growth factor in a weight ratio of 2:11.

[0031] Preparation:

[0032] S1 Add cell growth factors, taurine and fibronectin to the basal medium, stir for 20 minutes, add cholesterol, lipoic acid, chondroitin sulfate, biotin, insulin and myristic acid, continue stirring for 23 minutes, add peroxide hydrogenase, stirred for 20 minutes to obtain a mixture;

[0033] S2 Adjust the pH of the mixture obtained in step S1 to 6.7, filter and ...

Embodiment 2

[0034] Embodiment 2, a kind of medium for cultivating hepatocytes

[0035] The medium for cultivating hepatocytes includes basal medium and additives added in the basal medium, and the ingredients of the additives include: cholesterol 7mg / L, lipoic acid 0.4mg / L, cell Growth factor 35μg / L, chondroitin sulfate 0.5mg / L, biotin 16μM, insulin 1.3mg / L, catalase 17mg / L, taurine 0.3g / L, myristic acid 8mg / L, fibronectin 22μg / L.

[0036] The basal medium is RMPI 1640 medium.

[0037] The cell growth factor is composed of epidermal growth factor and hepatocyte growth factor in a weight ratio of 5:7.

[0038] Preparation:

[0039] S1 Add cell growth factors, taurine and fibronectin to the basal medium, stir for 30 minutes, add cholesterol, lipoic acid, chondroitin sulfate, biotin, insulin and myristic acid, continue stirring for 35 minutes, add peroxide hydrogenase, stirred for 20 minutes to obtain a mixture;

[0040] S2 Adjust the pH of the mixture obtained in step S1 to 7.3, filter...

Embodiment 3

[0041]Embodiment 3, a kind of medium for cultivating hepatocytes

[0042] The medium for cultivating hepatocytes includes basal medium and additives added in the basal medium, and the ingredients of the additives include: cholesterol 5mg / L, lipoic acid 0.3mg / L, cell Growth factor 28μg / L, chondroitin sulfate 0.4mg / L, biotin 15μM, insulin 1.1mg / L, catalase 14mg / L, taurine 0.2g / L, myristic acid 7mg / L, fibronectin 17μg / L.

[0043] The basal medium is RMPI 1640 medium.

[0044] The cell growth factor is composed of epidermal growth factor and hepatocyte growth factor in a weight ratio of 3:10.

[0045] Preparation:

[0046] S1 Add cell growth factors, taurine and fibronectin to the basal medium, stir for 23 minutes, add cholesterol, lipoic acid, chondroitin sulfate, biotin, insulin and myristic acid, continue stirring for 32 minutes, add peroxide hydrogenase, stirred for 20 minutes to obtain a mixture;

[0047] S2 Adjust the pH of the mixture obtained in step S1 to 6.9, filter...

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Abstract

The invention belongs to the technical field of cell culture media and particularly relates to a culture medium for culturing hepatocytes and a preparation method thereof. The culture medium for culturing hepatocytes comprises a basal culture medium and additives added into the basal culture medium, wherein the additives comprise the following components according to final concentration: 4-7mg / L of cholesterol, 0.2-0.4mg / L of lipoic acid, 20-35 micrograms / L of cell growth factors, 0.2-0.5mg / L of chondroitin sulfate, 13-16 micromoles / L of biotin, 0.9-1.3mg / L of insulin, 10-17mg / L of catalase, 0.1-0.3g / L of taurine, 6-8mg / L of myristic acid and 14-22 micrograms / L of fibronectin. The culture medium for culturing hepatocytes is free of fetal calf serum and can provide adequate nutrition and good environment needed for growth and proliferation of cells.

Description

technical field [0001] The invention belongs to the technical field of cell culture medium, and in particular relates to a culture medium for culturing hepatocytes and a preparation method thereof. Background technique [0002] As one of the most important organs in the human body, the liver undertakes various complex functions such as synthesis, secretion, metabolism, and detoxification. Once a large amount of necrosis of liver cells is caused by various reasons, liver failure will occur, leading to metabolic disorders and the accumulation of toxic substances, which in turn aggravate the damage of liver cells, affect the regeneration of liver cells, and then form liver failure vicious circle. Acute liver failure is a serious liver disease with a mortality rate as high as 60-90%. Currently, the most effective treatment for this is liver transplantation. However, due to lack of donor organs, high cost, long-term use of immunosuppressants and other reasons, patients often d...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/067C12N2501/11C12N2501/12C12N2501/33C12N2501/39C12N2501/71C12N2501/90C12N2501/998C12N2501/999
Inventor 叶宗耀
Owner 叶宗耀