Method for detecting cell injury repair ability, and special chip thereof

A cell and chip technology, applied in the field of detection of cell damage repair ability, method and its special chip, can solve the problems of large changes in the size and width of the wound area, difficult control of the initial damage area, and the efficiency of cell damage migration, etc., to achieve easy reversibility Sealing, biocompatibility, high stickiness effect on the bottom surface

Inactive Publication Date: 2009-11-25
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

The scratch method relies on manual operation, the size and width of the wound area vary greatly, the initial damage area is not easy to control, it is difficult to accurately locate the defect when taking pictures, and the scratch may cause damage to the surrounding cells and affect their migration efficiency. Therefore, the results of this "scratch" wound healing test are not ideal due to the multifactorial interaction between cells, which often leads to poor experimental repeatability. More importantly, in practical applications, this qualitative observation (rather than Quantitative measurement) is difficult to be applied to high-throughput research

Method used

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  • Method for detecting cell injury repair ability, and special chip thereof
  • Method for detecting cell injury repair ability, and special chip thereof
  • Method for detecting cell injury repair ability, and special chip thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0023] Will figure 1 The chip shown (there are 4 structural units in total, and each structural unit has 3 columnar structures, so 12 cell defect regions with the same size and shape can be generated at the same time) was inoculated with a high concentration (about 10 7 / ml) Human gastric mucosal epithelial cells (GES-1) were placed at 37°C, 5% CO 2 In the incubator overnight, after the cells adhered to the wall and the density was greater than 80%, the microfluidic chip was gently removed. The control group was added with high-sugar DMEM medium containing 12% newborn bovine serum, and the EGF group was added with 90ng / ml EGF and 12% newborn calf serum high-glucose DMEM medium, immediately look for the damaged area under the microscope and take pictures. After taking pictures, continue to place them in the incubator for cultivation, then take pictures every 8 hours, and observe continuously for 24 hours. It can be seen that EGF (90ng / ml) can significantly promote the damage ...

Embodiment 2

[0028] Will figure 1 The indicated chips were inoculated with high concentrations (approximately 10 7 / ml) Human lens epithelial cells cultured in vitro were placed at 37°C, 5% CO 2 After overnight in the incubator, after the cells adhered to the wall and the density was greater than 80%, the microfluidic chip was gently removed, and cells containing different concentrations of basic fibroblast growth factor (basic fibroblast growth factor, b-FGF) were added. culture medium, immediately look for damaged areas under a microscope and take pictures. After taking pictures, continue to place them in the incubator for cultivation, then take pictures every 8 hours, and observe continuously for 24 hours. It can be seen that 5μg / L bFGF can significantly promote the injury repair speed of human lens epithelial cells, and the promotion effect gradually increases with the increase of b-FGF concentration.

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Abstract

The invention provides a method for detecting cell injury repair ability and a special chip thereof. The chip consists of 2 to 99 identical cell culture structure units, wherein the structure units are connected with each other through a public cell sample pool, and each structure unit comprises a column-shaped structure, a baffle structure and a waste liquid pool. The invention creatively integrates cell culture, defect region formation, injury-repair process starting and other functional units on one chip, and adopts the function of the column-shaped structures in blocking cells to form cell defect regions with the same shape and size. Compared with the prior scratch method, the method has the advantages of simplifying artificial operation, providing exactly the same initial defect region, reducing cell consumption, having high flux and ensuring that the injury repair process is easy to photograph in fixed positions.

Description

technical field [0001] The invention relates to a technique for measuring cell damage repair, and in particular provides a method for detecting cell damage repair ability and a dedicated chip thereof. Background technique [0002] The environment in which organisms live is constantly changing, and there are many pathogenic factors such as bacteria, viruses, and allergens. The continuous cell layer is easily damaged under the continuous action of pathological factors, and cells fall off to form defects, and the tissue reconstruction and functional reconstruction process after damage depends on the damage repair ability of cells. The process of injury repair begins with the polarization of cells around the wound to the wound site and extension of synapses, and then the cells begin to migrate and proliferate to gradually heal the wound. The above process is similar whether in the whole tissue or at the level of individual cells. Damage repair experiments have been used as an ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/34C12Q1/02
Inventor 秦建华张敏林炳承李艳峰
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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