Complete culture medium for culturing hepatocytes

A technology of complete medium and hepatocytes, which is applied in the cultivation process, animal cells, tissue culture, etc., can solve the problems of discounting the effect of cytology experiments, and achieve the effects of avoiding ethical risks, saving costs, reducing time consumption and costs

Pending Publication Date: 2019-04-30
ZHUJIANG HOSPITAL SOUTHERN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Limited by this, in the process of cell culture and drug testing, it is difficult to restore the natural state of cells in the organism, so the effect of cytological experiments will be greatly reduced, and the metabolism of drugs and cells in the body cannot be simulated to the greatest extent

Method used

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  • Complete culture medium for culturing hepatocytes
  • Complete culture medium for culturing hepatocytes
  • Complete culture medium for culturing hepatocytes

Examples

Experimental program
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preparation example Construction

[0051] One, the preparation of described complete culture medium:

[0052] Only the Hyclone DMEM medium formula is selected as the basal medium in the examples under the specific implementation mode, and other basal mediums can be selected under the premise that other experiments or culture conditions permit and do not deviate from the idea of ​​the present invention. Described Hyclone DMEM medium formula is as follows:

[0053] Inorganic salts include at least:

[0054]

[0055] Amino acids include at least:

[0056]

[0057] Vitamins include at least:

[0058]

[0059]

[0060] Other substances are at least:

[0061] D-glucose 4500mg / L

[0062] Sodium phenol red 15.9mg / L

[0063] Sodium bicarbonate 3700mg / L

Embodiment 1

[0064] Embodiment 1: the preparation of complete medium 1:

[0065] In the present embodiment, the powder of the above-mentioned Hyclone DMEM medium formula is used as the base medium and the complete medium of the present invention is configured on this basis. The specific steps are as follows:

[0066] 1) Disinfect or sterilize the equipment and utensils used for configuration, including but not limited to serum bottles, magnetic stirring beads, volumetric flasks, beakers, pH meters, etc.;

[0067] 2) collecting the portal vein blood of fresh Tibetan minipigs;

[0068] 3) Centrifuge the portal vein blood with a centrifugal force of 3000×g for 12 minutes, and remove the supernatant;

[0069] 4) heating the lower supernatant at 56° C. to inactivate the complement to obtain the inactivated supernatant;

[0070] 5) Filter the inactivated supernatant through a microporous membrane with a filter hole diameter of 0.22 μm, collect porcine portal vein serum, and store it for future...

Embodiment 2

[0079] Embodiment 2: the preparation of complete medium 2:

[0080] In the present embodiment, the powder of the above-mentioned Hyclone DMEM medium formula is used as the base medium and the complete medium of the present invention is configured on this basis. The specific steps are as follows:

[0081] 1) Disinfect or sterilize the equipment and utensils used for configuration, including but not limited to serum bottles, magnetic stirring beads, volumetric flasks, beakers, pH meters, etc.;

[0082] 2) collecting the portal vein blood of fresh Tibetan minipigs;

[0083] 3) Centrifuge the portal vein blood with a centrifugal force of 3000×g for 12 minutes, and remove the supernatant;

[0084] 4) heating the lower supernatant at 56° C. to inactivate the complement to obtain the inactivated supernatant;

[0085] 5) Filter the inactivated supernatant through a microporous membrane with a filter hole diameter of 0.22 μm, collect porcine portal vein serum, and store it for future...

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Abstract

The invention provides a complete culture medium for culturing hepatocytes. Complete culture medium components comprise a basic culture medium and mixed serum, wherein the mixed serum is composed of portal vein serum and fetal bovine serum. The complete culture medium provided by the invention can be applied to culture of a hepatoma cell line HepG2, and can also enhance the expression of pigment metabolic enzymes in the hepatocytes. The gene expression level of metabolic enzymes in the hepatocytes is enhanced in the premise of ensuring the normal growth of the hepatocytes and the survival of the hepatocytes.

Description

technical field [0001] The invention relates to the field of cell culture, in particular to a complete culture medium for culturing hepatocytes and improving the expression of enzyme genes. Background technique [0002] In the process of drug development and drug testing, drug safety evaluation is an important link, including biochemical analysis, cytology experiments, toxicology experiments, animal experiments and human clinical experiments. If the metabolites in the body can be better simulated at the cellular level in the early stage, it will reduce the risk of failure in animal experiments and human clinical trials in the later stage. [0003] The main organ of drug metabolism in the body is the liver, and the smallest unit that performs metabolic functions is the liver cell. The study of drug metabolism in the body at the biochemical level mainly involves two processes: phase I metabolism and phase II metabolism. Specifically, phase I metabolism refers to that under t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/067C12N2500/84
Inventor 彭青高毅李海燕李阳梁康檐黎少翁骏
Owner ZHUJIANG HOSPITAL SOUTHERN MEDICAL UNIV
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