Novel coronavirus intestinal infection model construction method based on micro-fluidic chip

A microfluidic chip and coronavirus technology, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, and resistance to vector-borne diseases, etc., can solve the problem of difficult to reflect the dynamic cell biomechanical environment and restrict the infection of new coronaviruses Research on drug research and development, low accuracy of human response, etc.

Pending Publication Date: 2022-06-03
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the context of the current outbreak of new coronaviruses that urgently need specific drugs, the animal experiment cycle is long and expensive, and there are many restrictions on its wide application
However, static two-dimensional cell culture is completely different from the growth environment of cells in vivo, and it is difficult to reflect the dynamic cell biomechanical environment in vivo, and the accuracy of drug test results in predicting human response is low
Generally speaking, the existing drug evaluation research has some shortcomings in terms of methodology and technical means, which largely restrict the research and development of new coronavirus infection and drug research and development.

Method used

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  • Novel coronavirus intestinal infection model construction method based on micro-fluidic chip
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  • Novel coronavirus intestinal infection model construction method based on micro-fluidic chip

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Embodiment 1

[0031] Use the microfluidic chip designed and fabricated by the laboratory, see the configuration figure 1 . The microfluidic chip chip is formed by bonding and sealing the upper and lower layers of PDMS. The lower chip includes vascular endothelial cell inlet pool 1, vascular endothelial cell outlet pool 2, intestinal alveolar epithelial cell inlet pool 3, and intestinal alveolar epithelial cell outlet pool 4 , both the outlet and the inlet are connected to the perfusion tube;

[0032] In the microfluidic chip provided by the present invention, the two layers of PDMS channels of the microfluidic chip are separated by a PDMS porous membrane, and the height of the vascular endothelial cell culture chamber and the intestinal alveolar epithelial cell culture chamber is 200-1000 μm.

Embodiment 2

[0034] A method for constructing a novel coronavirus intestinal infection model based on microfluidic chip technology, using the above-mentioned microfluidic chip, according to the following steps:

[0035] Prepare 20-200 μg / ml type I collagen solution, add it into the inlet pool of intestinal epithelial and endothelial cells with a pipette, 30-100 μl per well; add 1 ml of PBS buffer to a petri dish, place it in an incubator for 10-48 h, Rinse 1-3 times with PBS; make a suspension of endothelial cells, add 30 μl of the suspension to the inlet pool of vascular endothelial cells, and quickly suck 10 μl of cell culture medium from the outlet pool to promote cells to quickly and uniformly under the action of gravity flow. into the cell culture chamber. When the cells in the cell culture chamber are evenly distributed, immediately turn the chip upside down and place it in a 37°C incubator; after standing upright for 2 hours, take it out and observe that the cells adhere closely to ...

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Abstract

The invention provides a novel coronavirus intestinal infection model construction method based on a micro-fluidic organ chip, and particularly aims at simulating and constructing a series of intestinal tissue pathophysiological changes such as intestinal barrier integrity damage, mucus secretion abnormity and intestinal immune response activation occurring after novel coronavirus infection. The micro-fluidic chip is mainly composed of two layers of micro-channels, the channels are separated by a polydimethylsiloxane wedge-shaped porous membrane, each layer of channel is provided with an inlet and an outlet, and a static or dynamic environment can be formed through fluid. By respectively inoculating intestinal epithelial cells and endothelial cells into the upper channel and the lower channel on the chip and introducing circulating immune cells into the lower channel after virus infection, the miniature intestinal tissue on the chip is formed. Novel coronavirus particles are added into an upper channel of the chip, so that novel coronavirus exposure of intestinal tissues is simulated. The invention provides a multi-factor participated tissue level research system for the research of the SARS-CoV-2 infected intestinal pathogenesis.

Description

technical field [0001] The invention relates to the technical field of applying microfluidic chip technology to intestinal physiology / pathology research, in particular to a method for constructing a novel coronavirus intestinal infection model based on microfluidic chip technology. Background technique [0002] SARS-CoV-2 is a severe acute respiratory syndrome-related coronavirus that is mainly transmitted from person to person through close contact and droplets produced by coughing or sneezing, mainly through interaction with the receptor angiotensin-converting enzyme. 2 (ACE2) binds into human cells. SARS-CoV-2 is mainly transmitted through the respiratory tract, causing flu-like symptoms. However, patients often present with gastrointestinal symptoms such as diarrhea, vomiting, and abdominal pain. In addition, SARS-CoV-2 receptors are abundantly expressed in intestinal epithelial cells, which may be one of the target organs for virus attack. Researchers have detected t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02C12Q1/6883C12Q1/6837G01N33/569G01N33/68
CPCG01N33/5005G01N33/56966G01N33/6893C12Q1/6883C12Q1/6837C12Q2600/158G01N2800/26Y02A50/30
Inventor 秦建华郭雅琼张敏王鹏
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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