Hepatic cell specificity culture matrix gel and preparation method thereof

A culture substrate and culture method technology, applied in cell culture support/coating, biochemical equipment and methods, tissue culture, etc., can solve the problems of inability to guarantee biological safety and difficulty in completely simulating the microenvironment of liver cells, and achieve The effect of maintaining function and differentiation, simple preparation method, and high cell activity

Pending Publication Date: 2018-10-16
ZHUJIANG HOSPITAL SOUTHERN MEDICAL UNIV
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  • Application Information

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Problems solved by technology

However, due to the specificity of the structural agent components in tissues and organs, it is difficult for existing matrix products to completely simulate the microenvironment required for the growth of hepatocytes in vivo, and some products such as Matrigel cannot guarantee their biological safety
Therefore, there is still a lack of a matrix product that can completely simulate the microenvironment of the growth of hepatocytes in vivo for the in vitro culture of hepatocytes

Method used

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  • Hepatic cell specificity culture matrix gel and preparation method thereof
  • Hepatic cell specificity culture matrix gel and preparation method thereof
  • Hepatic cell specificity culture matrix gel and preparation method thereof

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preparation example Construction

[0051] The preparation method of the liver cell-specific culture matrix gel of one embodiment, such as figure 1 As shown, at least the following steps are included:

[0052] Step S100: obtaining a decellularized liver scaffold, and making the decellularized liver scaffold into a dry powder;

[0053] Step S200: enzymatically hydrolyzing the dry powder with a pepsin solution to obtain an enzymatic hydrolyzate;

[0054] Step S300: Adjust the pH value and ion concentration of the enzymolysis solution, and form a gel after incubation.

[0055]In step S100, the preparation method of the decellularized liver scaffold includes the following steps:

[0056] Step S110: Obtain liver tissue slices, and wash the tissue slices with buffer solution containing EDTA. In this step, the liver tissue slices are taken from human or non-human (for example, from pigs) livers, and after the liver is sufficiently freed from the animal body, tissue slices are made. The size of the slices can be 1cm×...

Embodiment 1

[0067] The preparation method of the hepatocyte-specific culture matrix gel for hepatocyte culture comprises the following steps:

[0068] Preparation of the decellularized liver scaffold: the pig liver was made into tissue slices, and the size of the slices was 1 cm×1 cm×0.2 cm. Firstly, use PBS buffer solution containing 0.02% EDTA to wash blood stains and other dirt on the tissue sections. Put the cleaned tissue slices into a PBS solution containing 0.5% SDS and stir for 48 hours, then rinse them repeatedly with sterile deionized water for 24 hours to remove the detergent components on the scaffold, and finally slice the tissue after decellularization Freeze-dried and crushed into powder.

[0069] Preparation of hepatocyte-specific culture matrix gel: Enzymolyze the powdered decellularized liver scaffold obtained in the previous steps with an acidic pepsin solution with an optimal concentration and pH value, and the acidic pepsin solution with an optimal concentration and ...

Embodiment 2

[0072] The preparation method of the hepatocyte-specific culture matrix gel for hepatocyte culture comprises the following steps:

[0073] Preparation of the decellularized liver scaffold: After the liver from human or non-human source is sufficiently dissociated, tissue slices are made, and the size of the slices is 1 cm×1 cm×0.2 cm. Firstly, use PBS buffer solution containing 0.015% EDTA to wash away blood stains and other dirt on the tissue sections. The cleaned tissue slices were placed in PBS buffer solution containing 0.4% SDS and stirred for 48 hours, then washed repeatedly with sterile deionized water for 24 hours to remove the detergent components on the scaffold, and finally the decellularized tissue The slices were freeze-dried and pulverized into a powder by stirring.

[0074] Preparation of hepatocyte-specific culture matrix gel: Enzymolyze the powdered decellularized liver scaffold obtained in the previous steps with an acidic pepsin solution with an optimal con...

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Abstract

The invention discloses a preparation method of hepatic cell specificity culture matrix gel and the corresponding hepatic cell specificity culture matrix gel. The preparation method comprises the following steps that a decellularized liver support is obtained and made into dry powder; the dry powder is enzymatically hydrolyzed with a pepsin solution to obtain enzymatic hydrolysate; and the pH value and ionic concentration of the enzymatic hydrolysate are adjusted and the gel is formed after incubation. The preparation method of the hepatic cell specificity culture matrix gel is simple in process, and the hepatic cell specificity culture matrix gel is beneficial to the good growth of cells cultured in the hepatic cell specificity culture matrix gel.

Description

technical field [0001] The invention belongs to the technical field of in vitro cell culture, and in particular relates to a hepatocyte-specific culture matrix gel and a preparation method thereof. Background technique [0002] Hydrogels are increasingly used in tissue engineering because of their flexibility, porous properties, shape changeability and adjustable strength. Although the synthetic polymer gel has good property controllability, it is biologically inert and does not have the biological function of promoting the growth of cells and tissues in tissue engineering applications. Therefore, in tissue engineering applications, natural polymer gels that can provide bioactivity will be selected as the main component or active ingredient. [0003] So far, the only radical cure for end-stage liver disease is liver transplantation. However, the severe imbalance between supply and demand of liver sources makes most patients die while waiting for transplantation. Therefore,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/067C12N2533/90
Inventor 程远潘明新何国林曹玉伦李阳高毅
Owner ZHUJIANG HOSPITAL SOUTHERN MEDICAL UNIV
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