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Integrated microfluidic tissue chip and large-scale stimulant screening and analyzing method

A tissue chip and stimulant technology, which is applied in the field of microfluidic chips, can solve the problem of difficult to achieve high-throughput preparation of three-dimensional tissue-like tissue with a wide range of concentration gradients, and achieve simple positioning and recovery operations, and high-throughput operation and analysis results. Effect

Pending Publication Date: 2022-05-13
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing microfluidic chips are difficult to realize the generation of large-scale concentration gradients, the high-throughput preparation of three-dimensional tissue-like tissues, the simultaneous stimulation and analysis of large-scale concentration gradients of tissue-like tissues, and the possibility of tissue-like samples under different stimulation conditions. Recycling

Method used

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  • Integrated microfluidic tissue chip and large-scale stimulant screening and analyzing method
  • Integrated microfluidic tissue chip and large-scale stimulant screening and analyzing method
  • Integrated microfluidic tissue chip and large-scale stimulant screening and analyzing method

Examples

Experimental program
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Effect test

Embodiment 1

[0027] An integrated microfluidic chip used in this example was designed and manufactured by the applicant's laboratory. The material used to prepare the integrated microfluidic chip is polydimethylsiloxane, which is irreversibly sealed on the glass surface coated with polydimethylsiloxane.

[0028] Such as figure 1 , 2 as shown, figure 1 It is a structural schematic diagram of the present invention, which consists of a flow layer and a control layer. The fluidized layer of the chip includes two liquid inlets 1 and 2, nine outlets 3, one chemical concentration gradient generator 4, and nine microcavities 5. Among them, the chemical concentration gradient generator has 2 starting ports 21 and 22, 9 outlet ports 23, 1 non-irritant unit 24, 7 flow distribution and mixing units 25, and 1 stimulating unit 26, and each starting port is connected to One liquid inlet is correspondingly connected, and each outlet port is correspondingly connected with one microcavity 5; each microc...

Embodiment 2

[0041] This example provides the large-scale stimulus screening analysis of the integrated microfluidic tissue chip in Example 1.

[0042] First, use a microsyringe pump to inject a bovine serum albumin solution with a concentration of 2 mg / mL at a flow rate of 10 μL / min from the fluid inlets (1 and 2) of the chip fluid layer, and the bovine serum albumin enters all micropipes and microtubes in the fluid layer of the chip. cavity, incubate at 37°C for 3 hours, and perform anti-cell adhesion modification on the inner surface of all microchannels and microcavities;

[0043] Then perfuse with fresh cell culture fluid to clean all the micropipes and microcavities in the fluid layer of the chip;

[0044] Cell density was 5 x 10 5 Each / mL neural stem cell suspension is perfused into the microcavity through liquid inlet 1 and 2, such as image 3 shown;

[0045] Apply a certain air pressure to the aerodynamic microstructure to open the aerodynamic microstructure, such as Figure 4...

Embodiment 3

[0051] This example provides the large-scale stimulus screening analysis of the integrated microfluidic tissue chip in Example 1.

[0052] First, use a micro-syringe pump to inject a polyethylene glycol solution with a concentration of 1 mg / mL at a flow rate of 15 μL / min from the fluid inlets (1 and 2) of the chip fluid layer, and the polyethylene glycol enters all micropipes and microtubes in the fluid layer of the chip. cavity, incubate at room temperature for 2 hours, and perform anti-cell adhesion modification on the inner surface of all microchannels and microcavities;

[0053] Then perfuse with fresh cell culture fluid to clean all the micropipes and microcavities in the fluid layer of the chip;

[0054] Cell density was 1 x 10 6 Each / mL hepatic cell suspension is perfused into the microcavity from liquid inlets 1 and 2;

[0055] A certain air pressure (20psi) is applied to the pneumatic microstructure, the pneumatic microstructure is turned on, and the liver cells are...

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Abstract

The invention discloses an integrated microfluidic tissue chip and a large-scale stimulant screening analysis method. The chip is composed of a flow layer and a control layer. The flowing layer is provided with a liquid inlet, a chemical concentration gradient generator, a microcavity and an outlet. The chemical concentration gradient generator is provided with a starting end, a non-irritant unit, a flow distribution and mixing unit, an irritant unit and an outlet end, each starting end is correspondingly connected with a single liquid inlet, and each outlet end is correspondingly connected with a single microcavity; and each microcavity is correspondingly connected with a single outlet. And the control layer is provided with an air inlet, a pneumatic microstructure and an air inlet micro pipeline with a closed tail end. And all the pneumatic microstructures are communicated with the air inlet through micro pipelines with closed tail ends. Array cell localization, large-scale three-dimensional tissue-like preparation, large-scale order-of-magnitude concentration gradient type tissue-like stimulation and analysis thereof, and controllable recovery of different stimulated tissue-like samples can be realized in a single chip.

Description

technical field [0001] The invention relates to the technical field of microfluidic chips, in particular to an integrated microfluidic tissue chip and a method for screening and analyzing large-scale stimuli. It is used for arrayed cell positioning, large-scale three-dimensional tissue-like preparation, large-scale order of magnitude concentration gradient tissue-like stimulation and analysis, and controllable recovery of tissue-like samples with different stimuli. Background technique [0002] In vitro cell culture models are effective tools for basic biomedical research and drug development. In the traditional two-dimensional cell culture method, cells are usually cultured in culture dishes or well plates, and the environment in which the cells live is far from the growth environment of cells in tissues in vivo, and the functional expression and metabolic activities of cells cannot accurately reflect the real The in vivo state will affect the validity analysis of experime...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00C12M1/36C12M1/04C12N5/0797C12N5/071C12N5/09B01L3/00
CPCC12M23/16C12M27/00C12M29/00C12M33/04C12M41/34C12N5/0623C12N5/0671C12N5/0693C12N5/0631B01L3/5027C12N2513/00B01L2200/10
Inventor 刘文明孙美林张晋玮
Owner CENT SOUTH UNIV
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