Cell bidirectional invasion monitoring method based on micro-fluidic chip

A microfluidic chip and cell technology, applied in the field of cell biology research, can solve problems such as differences between different groups, affecting the accuracy of experimental data, etc.

Active Publication Date: 2017-04-19
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

At present, there have been preliminary studies on the use of microfluidic chips to monitor the process of cell invasion, but the existing microfluidic chip designs cannot achieve simultaneous observation and comparison. Parallel multi-group experiments in the research are likely to cause

Method used

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  • Cell bidirectional invasion monitoring method based on micro-fluidic chip
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  • Cell bidirectional invasion monitoring method based on micro-fluidic chip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Different effects of high glucose and low glucose on the invasion ability of U87 cells

[0042] After the 4 mg / mL collagen was poured into the collagen channel of the chip and coagulated, U87 was inoculated in the middle main channel 1, the left main channel 3 was added with low-glucose medium (glucose 5.5mM), and the right main channel 5 was added with high-glucose medium (glucose 5.5mM). 25mM), it can be observed that U87 has different selective invasion to the left collagen channel 2 and right collagen channel 4 under the action of high glucose and low glucose at the same time, and high glucose obviously promotes the invasion of U87 cells.

[0043] 24 hours after the cells were seeded on the chip, take a photo every 12 hours, and put the culture dish with the chip fixed into the stage incubator. Adjust the focal length and shooting parameters, and record the cell movement position and shape changes in real time when taking pictures with each microscope. A total of 10...

Embodiment 2

[0045] Different degrees of invasion induced by high glucose in U87-HIF and U87

[0046] After the 4mg / mL collagen was poured into the collagen channel of the chip and coagulated, U87 cells were inoculated in the left main channel 3, U87-HIF cells were inoculated in the right main channel 5, and high glucose medium (glucose 25mM) was added to the middle main channel 1. It was observed that U87 and U87-HIF invaded to different degrees on the left collagen channel 2 and right collagen channel 4 under the action of high glucose, and the invasion ability of U87-HIF cells was significantly stronger than that of U87 cells. 24 hours after the cells were seeded on the chip, the culture dish with the chip fixed was put into the stage incubator. Adjust the focal length and shooting parameters, and take photos under the microscope to record the cell movement position and shape changes in real time. The shooting took 108 hours in total, and the shooting results at the 48th hour are as fol...

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Abstract

The invention provides a cell bidirectional invasion monitoring method based on a micro-fluidic chip. According to the cell bidirectional invasion monitoring method, the micro-fluidic chip is adopted for monitoring the cell bidirectional invasion, the inducing or inhibiting effects of different cells or factors on the invasion of a target cell can be simultaneously observed and compared, and the inducing or inhibiting effects of one certain cell or factor on the invasion of different target cells also can be simultaneously observed. The micro-fluidic chip is mainly composed of three main channels and two collagen channels, wherein the three main channels used for cell culture horizontally communicate with the two collagen channels used for the cell migration observation; the left of the middle main channel (1) is connected with the left collagen channel (2), the right of the middle main channel (1) is connected with the right collagen channel (4), the left of the left collagen channel (2) is connected with the left main channel (3), and the right of the right collagen channel (4) is connected with the right main channel (5). The method has the feature of tracking the cell movement in real time, the accurate locating for the cell movement is realized, and meanwhile, the cell invasion capacity and selectivity can be observed and compared.

Description

technical field [0001] The invention relates to the technical field of applying microfluidic chip technology to real-time monitoring cell biology research, in particular to a microfluidic chip-based bidirectional cell invasion monitoring method. Background technique [0002] Since the development of cell biology, the main culture and research have relied on orifice plates and commercialized transwell chambers to focus on the observation of cell morphological changes, growth process, invasion and proliferation behaviors, etc. under the stimulation of single or multiple factors. Cell invasion generally starts from the cell's sensing of its microenvironmental stimuli. Microenvironmental stimuli activate a series of intracellular signal transduction pathways and gene transcription through cell surface receptors, and then through changes in cell polarity and cell adhesion. And de-adhesion, rearrangement of the cytoskeleton and other links, and finally complete the change of cell ...

Claims

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Application Information

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IPC IPC(8): C12Q1/02C12M3/00C12M1/34
Inventor 秦建华李中玉张旭
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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