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.
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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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