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Serum-free hepatocyte culture medium and preparation method thereof

A technology of hepatocytes and medium, applied in the field of serum-free medium for hepatocyte culture and its preparation, can solve problems affecting the accuracy and repeatability of research results and therapeutic effects, complex components, etc., to promote synthesis and cell growth and maintenance of cell viability

Inactive Publication Date: 2017-05-31
严志海
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the composition of serum is very complex, and the unknown substances and variable factors in it often affect the accuracy and repeatability of research results and treatment effects
In addition, FBS and other animal serum or serum protein and other animal-derived components will cause a variety of adverse reactions and diseases after entering the human body, laying a hidden danger for the clinical application of cell transplantation and BAL treatment

Method used

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  • Serum-free hepatocyte culture medium and preparation method thereof
  • Serum-free hepatocyte culture medium and preparation method thereof
  • Serum-free hepatocyte culture medium and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1 A kind of serum-free hepatocyte culture medium

[0028] A serum-free hepatocyte culture medium consisting of the following components and their concentrations: DMEM / F1235g / L, insulin 8.65mg / mL, transferrin 5.74mg / L, sodium selenite 8.2μg / L, epidermal growth factor 69μg / L, hepatocyte growth factor 78ng / mL, laminin 0.71μg / mL, dexamethasone 7.9mmol / L, oleic acid 0.112mmol / L, stearic acid 0.112mmol / L, palmitoleic acid 0.096mmol / L, Sodium bicarbonate 1.5g / L, tanshinone ⅡA 45μg / L and cryptotanshinone 4.3μg / L.

[0029] The preparation method of the serum-free hepatocyte culture medium of the present invention is divided into the following steps:

[0030] (1) Accurately weigh basal medium, insulin, transferrin, sodium selenite, epidermal growth factor, hepatocyte growth factor and laminin are dissolved in sterilized ultrapure water, stir evenly to obtain solution I ;

[0031] (2) Accurately weigh dexamethasone and dissolve it in 10 mL of absolute ethanol, then ...

Embodiment 2

[0035] Embodiment 2 A kind of serum-free hepatocyte culture medium

[0036] A serum-free hepatocyte culture medium consisting of the following components and their concentrations: DMEM / F1235g / L, insulin 8.65mg / mL, transferrin 5.74mg / L, sodium selenite 8.2μg / L, epidermal growth factor 69μg / L, hepatocyte growth factor 78ng / mL, laminin 0.71μg / mL, dexamethasone 7.9mmol / L, linoleic acid 0.064mmol / L, oleic acid 0.144mmol / L, palmitic acid 0.08mmol / L, Stearic acid 0.032mmol / L, sodium bicarbonate 1.5g / L, tanshinone ⅡA 45μg / L and cryptotanshinone 4.3μg / L.

[0037] The medium preparation method is similar to that of Example 1.

Embodiment 3

[0038] Embodiment 3 A kind of serum-free hepatocyte culture medium

[0039] A serum-free hepatocyte culture medium, consisting of the following components and their concentrations: DMEM / F1242g / L, insulin 1mg / mL, transferrin 1mg / L, sodium selenite 6μg / L, epidermal growth factor 10μg / L, Hepatocyte growth factor 15ng / mL, laminin 0.5μg / mL, dexamethasone 1mmol / L, oleic acid 0.035mmol / L, stearic acid 0.035mmol / L, palmitoleic acid 0.03mmol / L, sodium bicarbonate 1g / L, tanshinone ⅡA 10μg / L and cryptotanshinone 1μg / L.

[0040] The medium preparation method is similar to that of Example 1.

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Abstract

The invention relates to a culture medium for cell culture, and concretely relates to a serum-free hepatocyte culture medium and a preparation method thereof. The culture medium provided by the invention comprises a basic culture medium and additive components, wherein the additive components comprise insulin, transferrin, sodium selenite, epidermal growth factors, hepatocyte growth factors, laminin, dexamethasone, non-esterified fatty acid, sodium bicarbonate, tanshinone IIA and cryptotanshinone. The culture medium can promote the cell growth, and unexpectedly, the serum-free hepatocyte culture medium provided by the invention can greatly promote synthesis of albumin, and can maintain cell activity for a long time.

Description

technical field [0001] The invention relates to a medium for cell culture, in particular to a serum-free 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 die quickly due to disease ...

Claims

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Application Information

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IPC IPC(8): C12N5/071
Inventor 严志海
Owner 严志海
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