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Microfluidic chip and method for establishing in-vitro co-culture model of three kinds of cells

A technology of microfluidic chip and cell body, which is applied in tissue cell/virus culture devices, artificial cell constructs, animal cells, etc.

Active Publication Date: 2015-05-20
SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

AKH Achyuta et al simulated the neurovascular unit in the three-dimensional space of the microfluidic chip, in which the upper pool is the cell layer of brain microvascular endothelial cells, and the lower pool is the mixed culture of astrocytes, neurons and microglia, but the model exists The biggest defect is that the foot processes of astrocytes directly connect with the endothelial cell layer of brain microvascular

Method used

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  • Microfluidic chip and method for establishing in-vitro co-culture model of three kinds of cells
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  • Microfluidic chip and method for establishing in-vitro co-culture model of three kinds of cells

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Embodiment 1

[0058] figure 1 It is a schematic diagram of the structure of a microfluidic chip for establishing an in vitro co-culture model of three types of cells provided by the specific embodiment of the present invention. figure 2 yes figure 1 Partial enlarged view of the microfluidic chip. Such as figure 1 , 2 As shown in the present invention, a microfluidic chip for establishing the in vitro co-culture model of three types of cells includes: a first flow channel 1 in the middle, a symmetrical curved shape located on both sides of the first flow channel 1 The second flow channel 2 and the third flow channel 3 of the structure, and the thin flow channel 4 for connecting the first flow channel 1 and the second flow channel 2, the first flow channel 1 and the third flow channel 3;

[0059] The first channel 1 is used to inoculate astrocytes in the neurovascular unit;

[0060] The second flow channel 2 is used to inoculate neurons in the neurovascular unit;

[0061] The third cha...

Embodiment 2

[0066] image 3 It is a flow chart of a manufacturing method of a microfluidic chip for establishing an in vitro co-culture model of three types of cells provided by a specific embodiment of the present invention. Such as image 3 As shown, a method for manufacturing a microfluidic chip for establishing the in vitro co-culture model of three types of cells according to the present invention includes:

[0067] Step 1, fabricating thin flow channels 4 on the substrate material by photolithography;

[0068] Step 2, fabricating the first flow channel 1, the second flow channel 2 and the third flow channel 3 on the substrate material by photolithography;

[0069] Step 3, use the PDMS inverted mold to copy the photolithographic template structure;

[0070] Step 4, sealing with the glass substrate to form a closed flow channel, and completing the manufacture of the microfluidic chip structure.

[0071] A method for manufacturing a microfluidic chip used to establish an in vitro c...

Embodiment 3

[0091] Figure 4 It is a flow chart of a method for culturing cells using a microfluidic chip for establishing an in vitro co-cultivation model of three types of cells provided by a specific embodiment of the present invention. Such as Figure 4 As shown, a method for culturing cells using the above-mentioned microfluidic chip for establishing the in vitro co-culture model of the three types of cells according to the present invention includes:

[0092] Extraction and purification of neurons, astrocytes and brain microvascular endothelial cells;

[0093] After inoculating astrocytes in the first flow channel 1 of the microfluidic chip structure, inoculating neurons in the second flow channel 2, recording the growth status of the two, and labeling neurons with TJU1 and astrocytes with GFAP , observe the connection status of the two;

[0094] After the brain microvascular endothelial cells were inoculated in the third flow channel 3 of the microfluidic chip structure, astrocy...

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Abstract

The invention relates to the field of micromachining technology and tissue engineering technology, and particularly relates to a microfluidic chip and method for establishing an in-vitro co-culture model of three kinds of cells. The microfluidic chip is used for establishing a co-culture model of three kinds of cells; different cells are located in different areas through connected micro-channels in the microfluidic chip; and two kinds of cells can conduct cell communication through the connected micro-channels. Therefore, through the scheme, a new model capable of connecting astrocyte as a middle bridge in a neurovascular unit with the neuron and microvascular endothelial cell in a three-dimensional or plane space is established in vitro, a neurovascular network in the brain can be reproduced in vitro more scientifically and really, and a better platform is provided for clinical basic research on the central nervous system diseases and particularly the blood vessels of brain; and meanwhile, as an emerging biochip, the microfluidic chip is used for establishing the neurovascular in vitro, and thus the application range of the microfluidic chip is further widened.

Description

technical field [0001] The invention relates to the technical fields of microprocessing technology and tissue engineering, in particular to a microfluidic chip and a method for establishing three types of cells in vitro co-culture models. Background technique [0002] In recent years, with the increasing aging of my country's population and the influence of genetic and environmental factors, diseases of the central nervous system have gradually become an important challenge for clinical experts to face. For a long time, based on the research on the mechanism of neurodegenerative diseases, people have been using nerve cells as the target of action and a single target as the research target, trying to prevent the occurrence and development of neurodegenerative diseases with neuroprotective therapeutic strategies. However, for diseases of the nervous system, a single intracellular pathway or cell type is not enough, and effective therapeutic strategies must go beyond a single t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00C12N5/0793C12N5/079C12N5/071
Inventor 舒伟良陈艳
Owner SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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