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Preparation method for cell cultivation chip based on ITO glass substance and application thereof

A cell culture and glass substrate technology, applied in the field of glass cell culture chip preparation, can solve the problems of complex processing technology of hydrophobic glass material chips, and achieve the effect of removing CO2 supply, reducing complexity and maintaining stability

Inactive Publication Date: 2010-05-19
SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The object of the present invention is to provide a kind of cell culture chip preparation method based on ITO (Indium-Tin-Oxide, indium tin oxide) glass substrate and application thereof, overcome the hydrophobicity of existing polydimethylsiloxane (PDMS) chip Sexuality issues and the complex processing technology of glass chips provide good conditions for the stable cultivation of cells in vitro

Method used

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  • Preparation method for cell cultivation chip based on ITO glass substance and application thereof
  • Preparation method for cell cultivation chip based on ITO glass substance and application thereof
  • Preparation method for cell cultivation chip based on ITO glass substance and application thereof

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Experimental program
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Effect test

Embodiment 1

[0032] Cell culture glass chip manufacturing process such as figure 1 As shown, the details are as follows:

[0033] 1. To clean the ITO (Indium-Tin-Oxide, indium tin oxide) glass, first use acetone and alcohol to sonicate for 11 minutes, and then rinse it with deionized water;

[0034] 2. Spin-coat AZ4620 photoresist (2000 rpm, 30 seconds) on the side of the ITO glass where the ITO film is not sputtered, and bake at 80°C for 3 minutes;

[0035] 3. According to the designed mask (such as figure 2 shown) for exposure;

[0036] 4. Development, post-baking at 130°C, to obtain the graphic structure corresponding to the chip design;

[0037] 5. Mix polydimethylsiloxane (PDMS) monomer and curing agent (sylgard184 product provided by Dow Coming Company) in a weight ratio of 10:1, pour it on one side of the ITO film after degassing for 30 minutes, Curing at 80°C for 1 hour;

[0038] 6. Corrode the ITO glass in 1:6 diluted BOE etching solution for 50 minutes, transfer the pattern...

Embodiment 2

[0044] The 1640 medium was used as the culture medium, which was mixed with the L-15 medium at a ratio of 1:1, and supplemented with 10% fetal bovine serum and 100 Unit / ml streptomycin. Digest routinely cultured PIEC endothelial cells with trypsin digestion solution (containing 0.25% trypsin and 0.02% EDTA), centrifuge to remove the supernatant, disperse and resuspend the culture medium, and adjust the concentration of the cell suspension to 4×10 5 cells / mm3 , and then inhale the cell suspension with a syringe, and slowly inject it into the chip through a syringe pump. The cell-introduced chip is connected with a temperature control device and a culture fluid sampling device to carry out cell culture. The temperature control target was set at 37±0.5°C, and the injection speed of the culture solution was 8 μL / s. The chip is placed under an inverted microscope to observe the growth of cells in real time. Cell culture and observation of microsystems such as Figure 4 shown. ...

Embodiment 3

[0046] The culture medium adopts DMEM (high glucose) medium, which is mixed with L-15 medium at 1:1, and supplemented with 10% fetal bovine serum and 100 Unit / ml streptomycin. Digest conventionally cultured 3T3 cells with trypsin digestion solution (containing 0.25% trypsin and 0.02% EDTA), centrifuge to remove the supernatant, disperse and resuspend the culture medium, and adjust the concentration of the cell suspension to 4×10 5 cells / mm 3 ,, and then inhale the cell suspension with a syringe, and slowly inject it into the chip through a syringe pump. The cell-introduced chip is connected with a temperature control device and a culture fluid sampling device to carry out cell culture. The temperature control target was set at 37±0.5°C, and the injection speed of the culture solution was 8 μL / s. The chip is placed under an inverted microscope to observe the growth of cells in real time. Cell culture and observation of microsystems such as Figure 4 shown.

[0047] Figur...

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Abstract

The present invention relates to one kind of ITO glass substrate-based cell culture chip and its preparation process and application. The cell culture chip is prepared through the following steps: adopting ITO glass as substrate material, painting AZ4620 photoresist to the side without sputtered ITO film layer, exposing and developing; mixing PDMS monomer and curing agent, pouring the mixture to the side with ITO film and heating to cure; etching the ITO glass in etching solution; stripping PDMS from ITO film; bonding the PDMS film and ITO glass with etched micro pipeline under the action of oxygen plasma to obtain cell culture chip; and connecting external temperature control system to the side without bonded PDMS film of the ITO glass. The ITO glass substrate-based cell culture chip hasno hydrophobic problem, simplified preparation process and direct chip temperature control. It may be applied widely for cell research.

Description

technical field [0001] The invention belongs to the field of cell culture chips, in particular to a preparation method and application of a glass cell culture chip. Background technique [0002] Using CO 2 The conventional culture method of culturing cells in an incubator is a good method for studying living cells, and has been widely used in various fields such as virology, immunology, genetics, oncology, etc., but there are still obvious deficiencies. First of all, the cell operation method is cumbersome, it is difficult to control the microenvironment of cell growth, and it is difficult to fully simulate the real environment of cell life; and the number of cells required is huge, it is difficult to control and manipulate single cells, observation is not direct, and it takes centuries. The cell culture chip obtained by combining microfluidic chip and cell culture technology has obvious advantages. In terms of device size, the characteristic size of the chip can be compar...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C03C15/00C03C27/00
Inventor 吴蕾杨才表陈强赵辉赵建龙
Owner SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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