Culture solution for epatocyte scale culture

A technology of culture medium and liver cells, applied in tissue culture, biochemical equipment and methods, microorganisms, etc., can solve the problems of increasing lactic acid, affecting cell growth speed, inhibiting cell growth, etc., to reduce the production of H2O2 and enhance anti-oxidation effect, good calcium blocking effect

Active Publication Date: 2012-01-18
ZHUJIANG HOSPITAL SOUTHERN MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0008] 2. Relatively insufficient oxygen supply
This lack of oxygen can not only directly cause cell damage and affect the growth rate of cells, but also promote the anaerobic glycolysis of glucose in the medium, increase the production of lactic acid, make the pH value of the medium drop sharply, and affect the liver. Cells cause further damage
[0010] 3. Mechanical damage
However, under the action of many forces in the cell culture process (such as turbulent flow and shear force generated by air bubbles, and collision between microcarriers), cells attached to microcarriers will be damaged, inhibit cell growth, and cause cell death. die

Method used

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  • Culture solution for epatocyte scale culture
  • Culture solution for epatocyte scale culture
  • Culture solution for epatocyte scale culture

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Experimental program
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Effect test

Embodiment 1

[0044] Embodiment 1. Large-scale culture of human hepatocyte line CL-1 cells using the culture solution of the present invention; the group containing the protective agent is used as the experimental group, and the group without the protective agent is used as the control group;

[0045] 1. Preparation of culture medium:

[0046] Experimental group: Add 150ml of fetal bovine serum, 35mmol HEPES, 10000U of penicillin, 10000U of streptomycin and protective agent to each 1L of DMEN (high glucose type) medium, so that the concentration of each component in the protective agent and its concentration in the culture medium is: Salvia miltiorrhiza extract 60mg / L, astragalus extract 60mg / L, fructose 20mmol / L, vitamin C 30mg / L, insulin 10 -7 mmol / L.

[0047] Control group: 150ml of fetal bovine serum, 35mmol of HEPES, 10000U of penicillin and 10000U of streptomycin were added to each 1L of DMEN (high glucose type) medium.

[0048] 2. Microcarrier cytodex 3 pretreatment: put 0.25g micr...

Embodiment 2

[0060] Embodiment 2. Large-scale culture of human hepatocyte line CL-1 cells using the culture solution of the present invention; the group containing the protective agent was used as the experimental group, and the group without the protective agent was used as the control group;

[0061] 1. Preparation of culture medium:

[0062] Experimental group: Add 150ml of fetal bovine serum, 35mmol HEPES, 10000U of penicillin, 10000U of streptomycin and protective agent to each LDMEN (high glucose type) medium, so that the concentration of each component in the protective agent and its concentration in the culture medium is: Salvia miltiorrhiza extract 40mg / L, astragalus extract 40mg / L, fructose 10mmol / L, reduced glutathione 10mg / L, insulin 10 -8 mmol / L.

[0063] Control group: 150ml of fetal bovine serum, 35mmol of HEPES, 10000U of penicillin and 10000U of streptomycin were added to each 1L of DMEN (high glucose type) medium.

[0064] Microcarrier cytodex 3 pretreatment was carried...

Embodiment 3

[0068]Embodiment 3. Large-scale culture of human hepatocyte cell line CL-1 cells using the culture solution of the present invention; the group containing the protective agent was used as the experimental group, and the group without the protective agent was used as the control group;

[0069] 1. Preparation of culture medium:

[0070] Experimental group: Add 150ml of fetal bovine serum, 35mmol HEPES, 10000U of penicillin, 10000U of streptomycin and protective agent to each 1L of DMEN (high glucose type) medium, so that the concentration of each component in the protective agent and its concentration in the culture medium is: Salvia miltiorrhiza extract Food 100mg / L, astragalus extract 100mg / L, fructose 30mmol / L, vitamin C 10mg / L, reduced glutathione 50mg / L, insulin 10 -6 mmol / L.

[0071] Control group: add 150ml of fetal bovine serum, 35mmol of HEPES, 10000U of penicillin, and 10000U of streptomycin per 1 LDMEN (high glucose type) medium.

[0072] Microcarrier cytodex 3 pre...

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Abstract

The invention relates to a culture solution for epatocyte scale culture. The culture solution comprises culture medium and protecting agent, wherein the protecting agent comprises the following components with corresponding concentrations in the culture solution as follows: 40-100 mg / L of radix salviae miltiorrhizae extact, 40-100 mg / L of astragalus extract, 10-30 mmol / L of fructose, 10-50mg / L of anti-oxidant and 10<-8>-10<-6> mmol / L of insulin. The culture solution can adopt various mechanisms to effectively relieve the damage brought by various factors to hepatocytes in the process of epatocyte scale culture and inhibit the occurrence of hepatocellular apoptosis, thereby achieving the aims of further improving the density and function status of the epatocyte scale culture and effectively prolonging the time for the in-vitro survival of the hepatocytes.

Description

technical field [0001] The invention relates to a culture fluid for cell culture, in particular to a culture fluid for large-scale culture of hepatocytes. Background technique [0002] In the 1980s, with the maturity of hepatocyte isolation and culture technology and the development of biomedical engineering and other technologies, bioartificial livers based on cultured hepatocytes were gradually developed. People have always hoped to adopt this method of in vitro liver support. To replace the failed liver function of patients and create conditions for them to wait for liver transplantation or their own liver regeneration. At present, it is believed that at least 10 10 Cells of the above order of magnitude, and under the condition of ensuring the cell activity, the more the number of hepatocytes, the better the support and treatment effect. Therefore, the large-scale culture technology of hepatocytes in vitro has become the core technology of the development of bioartificia...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/08
Inventor 周焕城蒋泽生高毅张志刘勇
Owner ZHUJIANG HOSPITAL SOUTHERN MEDICAL UNIV
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