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Tumor cell migration dynamics monitoring method based on microfluidic chip

A microfluidic chip, tumor cell technology, applied in biochemical equipment and methods, microbial determination/inspection, enzymology/microbiology devices, etc., can solve problems such as being in the blank stage

Active Publication Date: 2012-10-24
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, the use of microfluidic chips to monitor the kinetics of tumor cell migration, especially the research and analysis of the process of tumor cell migration from a two-dimensional plane to a three-dimensional matrix is ​​still in a blank stage.

Method used

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  • Tumor cell migration dynamics monitoring method based on microfluidic chip
  • Tumor cell migration dynamics monitoring method based on microfluidic chip
  • Tumor cell migration dynamics monitoring method based on microfluidic chip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Cell Migration of HepG-2 Cells in Different Concentrations of Collagen

[0035] Three different concentrations of collagen working solutions were prepared, the concentrations were 1mg / mL, 2.5mg / mL, and 5mg / mL, and the distribution of collagen filaments with different concentrations was as follows: Figure 4 shown. Using the microfluidic chip designed and manufactured by the laboratory, the configuration is as follows: figure 1 shown. After perfusing the collagen into the chip and incubating for 30 minutes, a clear interface between the collagen and the two-dimensional plane can be seen ( figure 2 ). Add 10 μL of 1 x 10 from the cell inlet pool 6 / mL HepG-2 cell suspension, set the chip upright for 10 minutes, so that the cells can attach to the side of the collagen, such as image 3 shown. Take pictures to record the initial position of the cells, change the medium every 24 hours, and take pictures to record the cell movement. Counting the area of ​​HepG-2 cells...

Embodiment 2

[0037] Real-time monitoring of migration of SMMC-7721 in 2.5mg / mL collagen

[0038] Using the microfluidic chip designed and manufactured by the laboratory, the configuration is as follows: figure 1shown. After the 2.5mg / mL collagen was poured into the chip for coagulation, the same cell inoculation and culture methods as in Example 1 were adopted. 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, take a picture every 60 minutes, and record the cell movement position and shape changes in real time by taking pictures under the microscope. The shooting time is 19 hours. The shooting results are as follows: Figure 8 shown.

Embodiment 3

[0040] After adding relaxin D, SMMC-7721 migration movement analysis in 2.5mg / mL collagen.

[0041] Using the microfluidic chip designed and manufactured by the laboratory, the configuration is as follows: figure 1 shown. After the 2.5mg / mL collagen was poured into the chip for coagulation, the same cell inoculation and culture methods as in Example 1 were adopted. The relationship between the area of ​​SMMC-7721 cells migrating into collagen and the culture time is shown in Figure 9 shown. After the cells were seeded on the chip for 24 hours, 1 μg / mL relaxin D was added to the culture medium to stimulate the cells, and the morphological changes and migration behavior of the cells under the action of relaxin D for 24 hours were recorded. The results are as follows: Figure 10 As shown, the relationship between the migration area of ​​SMMC-7721 cells and the culture time is shown in Figure 11 shown.

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Abstract

The invention relates to a tumor cell migration dynamics monitoring method based on a microfluidic chip. The method particularly aims at the process of tumor cells migrating and moving into three-dimensional substrates from two-dimensional planes, the microfluidic chip mainly consists of a cell inlet pool (1), a collagen inlet pool (2), a waste liquid pool (3), cell culture rooms (4) and cell migration rooms (5), wherein the cell inlet pool (1) is connected with the upper parts of the cell culture rooms (5), the waste liquid pool (3) is connected with the lower parts of the cell culture rooms (5), and one cell culture room is connected with three cell migration rooms. The tumor cell migration dynamics monitoring method based on the microfluidic chip has the characteristic that the real-time tracking on the cell movement can be realized, and meanwhile, the accurate positioning at the beginning of the cell movement can be realized.

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 method for monitoring tumor cell migration dynamics based on a microfluidic chip. Background technique [0002] Since the development of cell biology, the main culture and research have relied on orifice plates and commercialized transwell chambers, focusing on the observation of cell morphological changes, growth processes, migration and proliferation behaviors, etc. under the stimulation of single or multiple factors. In many cell behavior studies, cell migration, as the basis of important life processes such as organism development and morphogenesis, has attracted much attention. Cell migration generally begins with the cell's response to its microenvironmental stimuli, which activate a series of intracellular signal transduction pathways and gene transcription through cell surface receptors, and the...

Claims

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Application Information

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IPC IPC(8): C12M1/34C12Q1/02
CPCC12M23/16C12M23/34C12M29/10C12M41/46
Inventor 秦建华马慧朋许慧高兴华
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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