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Method for guiding fixed-point cell growth by preparing chemical micro-patterns on surfaces of various materials

A micropattern and cell technology, which is applied in the field of preparation of highly selective cell micropatterns, can solve the problems of elastic stamp pollution, etc., and achieve the effects of low cost, mild preparation conditions and simple operation process

Inactive Publication Date: 2012-01-04
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the use of elastic stamps fundamentally limits the preparation of high-fidelity geometric patterns, and contamination by elastic stamp residues on the substrate often occurs.

Method used

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  • Method for guiding fixed-point cell growth by preparing chemical micro-patterns on surfaces of various materials
  • Method for guiding fixed-point cell growth by preparing chemical micro-patterns on surfaces of various materials

Examples

Experimental program
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Effect test

preparation example Construction

[0028] The preparation of the PEG self-assembled film in step 2) of the present invention adopts silanization reaction, the reaction adopts a condensation reflux device, the reaction time is 18h, and the reaction temperature is 60°C in a silicone oil bath. Flush the surface with copious amounts of ultrapure water. The preparation of PEGMA film is by atom transfer radical polymerization (ATRP), firstly, 3-aminopropyl triethoxysilane (APTES) is coupled on the surface, and then the surface initiator isobutyryl bromide is connected to the double Pyridine is used as a ligand, and cuprous bromide is used as a catalyst for surface polymerization reaction. The reaction atmosphere is nitrogen, and the reaction temperature is 25 degrees. After the reaction, the surface is washed with a large amount of ultrapure water. To prepare anti-protein adsorption membrane PEG on the surface of dopamine, put the dopamine-covered substrate into the PEG solution with amino groups at the end with a co...

example 1

[0034] Substrate surface hydroxylation: Place a circular glass slide with a diameter of 12 mm in piranha solution (H 2 SO 4 :H 2 o 2 =3:1v:v) Soak at 80°C for 30min until no air bubbles are generated, then sonicate in the order of ultrapure water, ethanol, and ultrapure water for 10min each, rinse with a large amount of ultrapure water after each sonication, and finally blow dry with nitrogen , baked in a drying oven at 120°C for 10 minutes to obtain a surface rich in silanol.

[0035] Preparation of PEG anti-protein adsorption film: silanization reaction is adopted, the concentration of PEG silane is 6mM, the reaction adopts condensation reflux device, the reaction time is 18h, the reaction temperature is 60°C in silicone oil bath, and toluene and isopropanol are used at the end of the reaction. Sonicate for 5 minutes, and rinse the surface with a large amount of ultrapure water after each sonication, and finally dry it with nitrogen.

[0036] UV light PEG anti-protein ad...

example 2

[0040] Except for the preparation of the anti-protein adsorption membrane, other steps are the same as Example 1. The preparation of PEGMA anti-protein adsorption membrane is by atom transfer radical polymerization (ATRP). First, triaminotriethoxysilane (APTES) is coupled on the surface rich in silicon hydroxyl groups, and then connected to the surface initiator isobutyryl bromide. , using bipyridine as a ligand and cuprous bromide as a catalyst for surface polymerization, the reaction atmosphere is nitrogen, the reaction temperature is 25 degrees, and the surface is washed with a large amount of ultrapure water after the reaction. The micropattern surfaces formed on polyethylene glycol PEG film and polyethylene glycol methacrylate PEGMA film were seeded at a concentration of 2 × 10 4 3T3 fibroblasts in cells / ml, the result of direct observation with a microscope. The continuous cell culture showed that the stability of the cell pattern on the PEGMA membrane can reach more th...

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Abstract

The invention discloses a method for guiding fixed-point cell growth by preparing chemical micro-patterns on the surfaces of various materials. The preparation process comprises the following steps of: 1) hydroxylating the surfaces of a glass sheet, a quartz sheet, a silicon sheet, a transition metal oxide and the like and coating dopamine on the surface of polymer or other inorganic materials; 2) obtaining dense anti-protein adsorption films on the pretreated surfaces by molecular self-assembly or an atom transfer radical polymerization method, wherein the dense anti-protein adsorption filmsare polyethylene glycol films or polyethylene glycol methacrylate films; and 3) covering mask plates on the anti-protein adsorption films, performing ultraviolet irradiation and adding fibronectin toobtain protein micro-patterns; and inoculating cells to obtain the cell patterns. The method has the advantages of simple operation, low cost, mild preparation conditions and no need of super clean experimental conditions. By adopting the method, stable and high-selectivity cell micro-patterns can be obtained on the surfaces of various materials; and the method has a broad application prospect inthe fields of basic researches on cell biology, tissue engineering, drug screening and cell-based biosensors.

Description

technical field [0001] The invention relates to the preparation of a stable and highly selective cell micropattern. Using molecular self-assembly technology combined with ultraviolet light, chemical micropatterns are prepared on the surface of various materials to guide the growth of cells at a fixed point. By adjusting the shape and size of micropatterns, cell adhesion, migration, differentiation and intercellular interactions can be controlled. , has broad application prospects in the fields of basic research of cell biology, tissue engineering, drug screening and cell-based biosensors. Background technique [0002] Cell patterning is an effective experimental method to study and control cell behavior and is widely used in various research fields such as cell biology, tissue engineering, drug screening, and wound therapy. There are a variety of techniques to achieve substrate patterning, including two categories: photolithography and soft lithography, through which patter...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08J7/12C03C17/30C04B41/49G03F7/00
Inventor 黄宁平余海黄刚汪燕艳吕兰欣
Owner SOUTHEAST UNIV
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