Study on in vivo fluid shearing force simulation cell behaviors on basis of microfluidic chip system

A microfluidic chip, the technology of fluid shear force, applied in the measurement/inspection of microorganisms, the cultivation device of tissue cells/viruses, the method of stress-stimulating the growth of microorganisms, etc. It can achieve the effect of low consumption of reagents and cells, easy observation and detection, and simple operation.

Inactive Publication Date: 2012-11-21
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Simulating the cell microenvironment, especially simulating the microenvironment related to cell biophysical factors such as fluid shear force, traction force, matrix physical properties, nano-surface and so on, relies on the existing technology, most of which have complex and huge equipment, cumbersome operation, and little impact on the microenvironment. The simulation is simple and cannot achieve real-time observation of cells. With the deepening of microfluidic chip research, the microfluidic chip system may become a new platform technology for bionic simulation of the cell microenvironment. Provide technical and theoretical support for tissue repair, reconstruction transplantation and bionics related to cell microenvironment research

Method used

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  • Study on in vivo fluid shearing force simulation cell behaviors on basis of microfluidic chip system
  • Study on in vivo fluid shearing force simulation cell behaviors on basis of microfluidic chip system
  • Study on in vivo fluid shearing force simulation cell behaviors on basis of microfluidic chip system

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Experimental program
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Effect test

Embodiment 1

[0032] Using the microfluidic chip system designed and produced by the laboratory, the configuration is as follows: figure 1 As shown, the depth of the channel is 50 μm, and the width ranges from 50 μm to 1 mm. A total of 4 different fluid shear forces can be generated. When the flow rate of the syringe pump is 9 μL / h, the fluid shear force is theoretically calculated. For: 7.7×10 -3 ,9.9×10 -4 ,1.4×10 -4 and2.9×10-5 dyne / cm 2 . Chip inoculation of mouse embryonic mesenchymal stem cells, inoculation density 1×10 5 cells / ML, after 24 hours of cell attachment, replace with α-MEM medium containing 15% fetal bovine serum, apply fluid shear force, and after 7 days, osteoblasts were stained with ALP, photographed under a microscope, and analyzed by Image Pro software. The result is as image 3 As shown, the osteogenic differentiation behavior of mesenchymal stem cells is different under different fluid shear forces, and it can be seen that 7.7×10 -3 dyne / cm 2 Osteogenesis wa...

Embodiment 2

[0034] Using the microfluidic chip system designed and produced by the laboratory, the configuration is as follows: figure 1 As shown, the depth of the channel is 50 μm, and the width ranges from 50 μm to 1 mm. A total of 4 different fluid shear forces can be generated. When the flow rate of the syringe pump is 9 μL / h, the fluid shear force is theoretically calculated. For: 7.7×10 -3 ,9.9×10 -4 ,1.4×10 -4 and2.9×10 -5 dyne / cm 2 . Chip inoculation of mouse embryonic mesenchymal stem cells, inoculation density 1×10 5 cells / ML, after 24 hours of cell attachment, replace with LG-DMEM+10%FBS+0.1μM dexamethasone+50μg / ml ascorbic acid+100mMβ-glycerophosphate sodium osteogenic differentiation medium, apply fluid shear force, 7 Days later, osteogenesis was stained with ALP, and after 14 days, osteogenesis was stained with OPN, photographed under a microscope, and analyzed by Image Pro software. The osteogenic differentiation behavior of mesenchymal stem cells is different under ...

Embodiment 3

[0036] Osteoprogenitor cells: using the microfluidic chip system designed and produced by the laboratory, the fluid shear force range is 7.7×10 -3 ,9.9×10 -4 ,1.4×10 -4 and2.9×10 -5 dyne / cm 2 . The chip was inoculated with bone progenitor cell line 3T3-E1 at a seeding density of 1×10 5 cells / ML, after 24 hours of cell attachment, the medium was changed every day, and fluid shear force stimulation was applied. After 7 days of mechanical stimulation, ALP, a characteristic marker of osteogenesis, was detected, and it was found that fluid shear force can promote osteogenesis of osteoprogenitor cells. direction differentiation.

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Abstract

The invention relates to a study on in vivo fluid shearing force simulation cell behaviors on the basis of a microfluidic chip system. An external precise fluid injection pump is used as a motive power source, a microfluidic chip soft etching technology is used as the basis, and the fluid resistance principle is utilized for designing and manufacturing a microfluidic passage resistance network, so cells in a microfluidic chip cell culture microchamber can feel fluid shearing force with different intensities, and further, the cell behavior change is studied. Through the microfluidic chip system, the process of the inoculation and long-term culture of cells, the fluid shearing force stimulation on the cells and the cell dyeing and analysis and detection are integrated on a functionalization chip to be completed, and the cell behavior analysis and the in vivo ultra-low fluid shearing force simulation on a microfluidic chip platform are realized.

Description

technical field [0001] The invention belongs to the field of microfluidic chip systems, and in particular relates to a study on the behavior of cells under simulated fluid shear force in vivo based on the microfluidic chip system. Background technique [0002] As an important science and technology of this century, microfluidic chip laboratory has demonstrated its unique advantages in many fields including biology, medicine, chemistry, materials science, etc. It has become an important platform for a new generation of cell research due to its characteristics of similarity, fast heat and mass transfer, and high throughput that can be integrated. With the continuous development in the past two decades, cell research based on the microfluidic chip system has made great breakthroughs, and related cell operations, such as cell culture, sorting, identification, etc., can basically be realized on the chip. Based on this, further cytological research, especially the research on cel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00C12M1/00C12M1/34C12Q1/02
CPCC12M41/00C12M35/04C12M23/16
Inventor 秦建华高兴华
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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