A microfluidic chip for screening antitumor drugs at the single sphere level and its application
An anti-tumor drug, microfluidic chip technology, applied in laboratory containers, microbial determination/inspection, biochemical equipment and methods, etc., can solve the problems of high price and complicated operation steps, and achieve simple operation , environment-friendly, and low experimental cost
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Embodiment 1
[0039] The SU-8 polymer template is made by photolithography technology, and the template is 12 groups of parallel microholes with 7 different diameters, the diameters are 200, 300, 400, 500, 600, 700, and 800 microns. 6 groups of the same size in parallel. Use conventional photoetching technology to make SU-8 template: apply SU-8 photoresist on a clean glass sheet, bake it at 95 degrees Celsius for 1 hour, remove the organic reagent, and wait for the SU-8 photoresist to cool ; Put the designed mask on and expose to UV for 90 seconds, then bake at 95 degrees Celsius for 15 minutes. Different from the conventional method, the development time of the SU-8 template of the present invention is 5 minutes, which is incomplete development. After drying the developer, it is melted at 85°C for 5 minutes, so that the structure of the micropores in the SU-8 template is changed from The right angle becomes an arc; after cooling down, the SU-8 template is fully exposed to the ultraviolet ...
Embodiment 2
[0041] Microsphere culture of U87 cells on a sealed PDMS polymer chip such as Figure 4 shown. The PDMS polymer chip was sterilized first, irradiated with ultraviolet light overnight, and then used 0.2% PF-127 solution for surface modification for 4 hours, washed twice with sterile water and PBS solution, and inoculated with U87 cell suspension. Under the action of gravity, it will settle to the bottom of the small hole on the surface of the chip and spontaneously form a three-dimensional cell sphere. As a result of designing microwells of different sizes, U87 cells formed multicellular microspheres of different sizes.
Embodiment 3
[0043] Screening of anti-tumor drugs at the single-ball level Microfluidic chips are used for the screening of anti-cancer drugs in U87 cells. Figure 5 It shows that after U87 cells were cultured for four days on PDMS polymer chips with concave holes of different sizes, 200 μmol / L resveratrol drug was added through the upper flow channel layer to act on U87 cell microspheres, and some multicellular microspheres The diameter of the cells became smaller, and the cell viability decreased, but the cell apoptosis did not appear. Figure 6 It shows that after U87 cells were cultured on PDMS polymer chips with different sizes of concave holes for four days, 1.5 μmol / L PI-103 drug was added through the upper flow layer, the diameter of some multicellular microspheres became smaller, and the cell viability decreased, cell apoptosis occurred.
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