Temperature-sensitive type cell culture surface and preparation method thereof

A temperature-sensitive, cell culture technology, applied in the field of cell culture, can solve the problems of inability to guarantee the automatic shedding effect of cells, unstable temperature-sensitive response function, high cost of required raw materials, etc. The effect of reducing production costs

Active Publication Date: 2016-01-06
GUANGZHOU JET BIOFILTRATION CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In the existing methods for preparing temperature-sensitive cell culture surfaces, some methods are used to physically coat temperature-sensitive compounds directly on the surface of cell culture devices. The temperature-sensitive response function achieved by this method is unstable. Sensitive compounds will fall off, so the effect of automatic cell detachment cannot be guaranteed; most of the others use high-energy electron beams to initiate the grafting of temperature-sensitive compounds on the cell culture surface, and a small number of them use ultraviolet light irradiation to initiate grafting, or plasma-induced grafting , or atom transfer radicals to initiate grafting, in these ways of realizing the grafting of surface temperature-sensitive compounds, some production equipment is expensive (such as high-energy electron beam, plasma), and some processes are complex and the cost of required raw materials is high, which is not conducive to the industry. (such as atom transfer radical initiation, ultraviolet light irradiation initiation), and some grafting efficiency is low, serious environmental pollution (such as plasma)

Method used

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  • Temperature-sensitive type cell culture surface and preparation method thereof
  • Temperature-sensitive type cell culture surface and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Weigh 0.5g of N-isopropylacrylamide monomer, and dissolve it in a mixed solvent of 7.5g of isopropanol and ethyl acetate with an azo initiator with a molar ratio of 5:100; The mixture solution is atomized into particles with a particle size of 2-1000 microns, and then the surface of the cell culture plate of the polystyrene material to be treated is exposed to the atomized steam atmosphere for 50 minutes to condense the mist on the surface to be treated, and finally Place it in an oven filled with nitrogen, take it out after incubating at 50-120°C for 50 minutes, and obtain a cell culture plate with a temperature-sensitive cell culture surface, with a grafting rate of 2.2 μg / cm 2 .

Embodiment 2

[0031] Weigh 0.7g of N-isopropylmethacrylamide monomer, and dissolve it in 10g of ethanol, propyl formate and dichloromethane in a mixed solvent of 10g of ethanol, propyl formate, and dichloromethane, and then use high-pressure Spray the temperature-sensitive mixture solution evenly on the surface of the polypropylene cell culture dish, and finally place it in an oven filled with helium, take it out after incubating at 90-120°C for 20 minutes, and obtain a temperature-sensitive cell culture dish The cell culture dish on the surface, the grafting rate is 1.7μg / cm 2 .

Embodiment 3

[0033] Weigh 0.6g of N-isopropylacrylamide monomer, dissolve in 6g of isopropanol, butyl acetate and dichloromethane mixed solvent with an azo initiator with a molar ratio of 30:100 to the monomer, and then spray it with air Spray the mixture solution on the surface of the cell culture flask made of polystyrene material, then place it in an air-filled oven, keep it at 50-120°C for 30 minutes, and then take it out to obtain a cell culture with a temperature-sensitive cell culture surface bottle, grafting rate 1.5μg / cm 2 .

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Abstract

The invention relates to a temperature-sensitive type cell culture surface and a preparation method thereof. The preparation method comprises the following steps: (1) preparing a temperature-sensitive primary liquid: adding temperature-sensitive compounds and radical initiators to solvents and stirring to dissolve the materials to obtain the temperature-sensitive primary liquid, wherein the mass ratio of the temperature-sensitive compounds to the solvents is (2-50) to 100; the mole ratio of the radical initiators to the temperature-sensitive compounds is (0.5-50) to 100; (2) reacting a cell culture surface for 5-120 minutes under the condition of 50-150 DEG C after distributing the temperature-sensitive primary liquid on the cell culture surface, thus obtaining the temperature-sensitive type cell culture surface. The preparation method has the beneficial effects that the cell culture surface with the temperature-sensitive response function can be obtained by directly initiating graft polymerization of the temperature-sensitive compounds on the cell culture surface by utilizing the common chemical radical initiators; compared with the prior art, the preparation method has the advantages that the production cost of the temperature-sensitive type cell culture surface is greatly reduced; the grafting ratio is high; the preparation method is environmentally friendly.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a temperature-sensitive cell culture surface and a preparation method thereof. Background technique [0002] In recent years, the technology of tissue regeneration engineering, biomedical engineering, and cell therapy industry has developed rapidly. It has become the current cell clinical practice to obtain a large number of cells, cell extracellular secretions, and cell therapy products through large-scale culture of animal or human cells in vitro. Important direction of application. For example, tissue trauma, skin cell culture in burn treatment and acquisition of new skin, tumor cell therapy, cell beauty, cell weight loss, etc., all require in vitro culture of target cells. [0003] Cells are mainly divided into suspension cells and adherent cells according to their in vitro culture characteristics. Suspension cells are suspended in the culture medium in the culture devi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00
CPCC12M23/20C12M33/00C12M3/04C08F257/02C08F220/56C08F112/08C08J7/12C08F293/00C08L25/06C08J2325/06
Inventor 袁建华陈勇李慧伦方想元
Owner GUANGZHOU JET BIOFILTRATION CO LTD
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