Two-layer cell culture system organ chip and preparation method thereof
A cell culture and organ-on-a-chip technology, applied in the field of biomedicine, can solve problems such as difficulty in miniaturization and integration, single use, and time-consuming, and achieve the effects of high parallel culture capacity, small dosage, and simple equipment
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Embodiment 1
[0028] The preparation of embodiment 1 chip
[0029] 1) Preparation of polymethacrylic acid (PMMA) chips
[0030] Computer-aided design software was used to design and draw the microstructure and microchannel pattern of the three-layer chip of the two-layer cell culture organ chip, and the microstructure and microchannel of the three-layer polymethacrylic acid (PMMA) chip were fabricated by numerical control CNC system. Wash each layer of chips with tap water and distilled water respectively, and wipe the fingerprints, oil stains and other stains remaining on the chip surface with ethanol. On the double-sided adhesive film, the microstructures and microchannels required for the preparation of the chip are processed by a lettering machine, and the three-layer chip is aligned, glued, and pressure-stitched to make four inlets, four cell culture chambers, and two upper and lower layers. A co-culture system, a detection cell with two layers of dialysis membranes, and a two-layer c...
Embodiment 2
[0033] Example 2 Preparation of microporous membrane two-layer co-culture system chip
[0034] Use computer-aided design software to design and draw the microstructure and microchannel patterns of the upper and lower layers of the two-layer cell culture organ chip, and use the numerical control CNC system to process and prepare the microstructure and microchannels of the three-layer polymethacrylic acid (PMMA) chip or Preparation of PDMS replication reverse template. Add a layer of photoresist microporous film in the cell culture area, the diameter of the micropore is 5μm ~ 20μm, and then align the two layers of chips, pressurized, or thermally cross-linked to synthesize the chip to make micropores Membrane two-layer cell culture organ chip.
Embodiment 3
[0035] Example 3 Parallel implantation and co-culture of cells
[0036] Close the drug inlet and outlet, inject different cells into the upper and lower cell solution inlet and outlet, control the flow rate of the syringe pump and the liquid difference height between the inlet and outlet, thereby controlling the laminar flow speed, so that the cell solution slowly flows through the microchannel into the cell culture chamber, Thus, the implantation of cells is completed, and the plate adherent culture or the three-dimensional gel matrix culture is carried out. The cell culture medium is supplied, and the cells are cultured for a period of time. After adhering or fixed growth, the medium is circulated and flowed to promote the stable growth and proliferation of cells, and maintain the vitality and function of cells.
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