Temperature-sensitive hepatic cell culture support material and preparation method thereof

A temperature-sensitive, cultured scaffold technology, applied in the field of intelligent tissue engineering scaffold materials, can solve the problems of loss of cell activity in the central part of the structure, inability to simulate the body tissue well, and slow discharge of nutrient metabolic wastes, etc. Compatibility, easy reaction control, wide range of effects

Inactive Publication Date: 2010-12-15
TIANJIN POLYTECHNIC UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Although many meaningful research results have been obtained on liver tissue engineering scaffold materials, according to traditional tissue engineering methods, when cells are cultured and expanded in vitro to form cell sheets or tissues, they must be transplanted into the main body together with scaffold materials, which will cause Many complicated problems are caused: (1) After the scaffold material is degraded in vivo, the space previously occupied by it will be filled with extracellular matrix, so that the cells cannot be tightly aggregated. The liver is an organ where cells are tightly aggregated, so that the transplantation The growth of the cells in the body cannot simulate the body tissue well; (2) For larger structures, around the scaffold material, the transplanted cells will grow healthily and are

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] (1) Surface cleaning: Wash polystyrene cell culture plate (TCPS) samples cut into 1cm×1cm with absolute ethanol to remove surface impurities, then rinse with distilled water several times, and vacuum dry at 60°C for later use.

[0035] (2) Temperature-sensitive carboxylation modification on the surface of TCPS: Dissolve 0.01mol N-isopropylacrylamide (NIPAAm) and 0.01mol acrylic acid (AAc) in 6.8mL dimethyl sulfoxide (DMSO), then add 0.001mol anthracene Sodium quinone-2-sulfonate (AQS) was used as a photosensitizer, and a few drops of isopropanol were added. Put the processed TCPS sample into the reaction solution, pass N 2 Seal for 15 minutes, irradiate with a high-pressure mercury lamp (λ=360nm, 1000W) for 0.5h, the distance between the high-pressure mercury lamp and the sample is 40cm, and the depth of the reaction solution is 1cm. Unreacted small molecules and unmodified polymers were then vacuum-dried for 1 day to obtain TCPS-NIPA-AAc on the thermosensitive carbox...

Embodiment 2

[0039] (1) surface cleaning: with example 1 (1).

[0040] (2) Temperature-sensitive carboxylation modification on the surface of TCPS: Dissolve 0.01mol N-isopropylacrylamide (NIPAAm) and 0.001mol acrylic acid (AAc) in 6.8mL dimethyl sulfoxide (DMSO), then add 0.001mol anthracene Sodium quinone-2-sulfonate (AQS) was used as a photosensitizer, and a few drops of isopropanol were added. Put the processed TCPS sample into the reaction solution, pass N 2 Seal for 15 minutes, and irradiate for 3 hours with a high-pressure mercury lamp (λ=360nm, 1000W). The distance between the high-pressure mercury lamp and the sample is 40cm, and the depth of the reaction solution is 1cm. The reacted small molecule and unmodified polymer were then vacuum-dried for 1 day to obtain TCPS-NIPA-AAc on the thermosensitive carboxylated surface of TCPS.

[0041] (3) Preparation of lactobionic acid (L-NH 2 ): mix 3g lactobionic acid (LA) and 18g ethylenediamine (NH 2 CH 2 CH 2 NH 2 ) were respective...

Embodiment 3

[0044] (1) surface cleaning: with example 1 (1).

[0045] (2) Temperature-sensitive carboxylation modification on the surface of TCPS: Dissolve 0.01mol N-isopropylacrylamide (NIPAAm) and 0.005mol acrylic acid (AAc) in 6.8mL dimethyl sulfoxide (DMSO), then add 0.001mol anthracene Sodium quinone-2-sulfonate (AQS) was used as a photosensitizer, and a few drops of isopropanol were added. Put the processed TCPS sample into the reaction solution, pass N 2 Seal for 15 minutes, irradiate with a high-pressure mercury lamp (λ=360nm, 1000W) for 0.5h, the distance between the high-pressure mercury lamp and the sample is 40cm, and the depth of the reaction solution is 1cm. Unreacted small molecules and unmodified polymers were then vacuum-dried for 1 day to obtain TCPS-NIPA-AAc on the thermosensitive carboxylated surface of TCPS.

[0046] (3) Preparation of lactobionic acid (L-NH 2 ): mix 2g lactobionic acid (LA) and 8g ethylenediamine (NH 2 CH 2 CH 2 NH 2 ) were respectively dissolv...

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Abstract

The invention discloses a temperature-sensitive hepatic cell culture support material and a preparation method thereof, and relates to an intelligent tissue engineering support material and technology, in particular to a temperature-sensitive hepatic cell culture support material, a method and technology. The support material is characterized in that: the surface of the material is relatively lyophobic at the temperature of 37 DEG C and is suitable for cell adherence/spreading and cell propagation, but the surface is hydrophilic at the temperature of 32 DEG C, so cells are desorbed from the surface automatically. The preparation method of the cell culture support material comprises the following steps of: (1) surface cleaning; (2) the temperature-sensitive carboxylation modification of a TCPS surface; (3) the preparation of amido lactobionic acid (L-NH2); and (4) the saccharifying modification of the TCPS surface. Through the preparation method, the biocompatibility and cell-bound loci of the support material are enhanced, the adsorption rate of the cells on the material surface is increased, and the intelligent support material which is suitable for hepatic tissue engineering is obtained.

Description

technical field [0001] The invention relates to an intelligent tissue engineering scaffold material and technology, in particular to a temperature-sensitive liver cell culture scaffold material and method technology. Background technique [0002] my country is a high-incidence area of ​​liver disease, among which severe hepatitis / liver failure is one of the liver diseases with the most serious condition and the highest mortality rate. The three common methods of treatment for severe hepatitis / liver failure are medical treatment, liver transplantation and artificial substitute treatment. Due to the lack of liver donors and the high cost of surgery, liver transplantation is difficult to be widely used in my country, and artificial substitutes cannot completely replace all functions of human organs and cannot automatically adjust according to the requirements of the human body, so they cannot cure diseases , can only temporarily relieve symptoms or prolong the patient's life in...

Claims

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Application Information

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IPC IPC(8): C12N5/071
Inventor 陈莉贺晓凌聂萍萍赵义平
Owner TIANJIN POLYTECHNIC UNIV
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