Microfluidic chip based on nanometer drug delivery system screening 3D solid tumor model and preparation method and application thereof
A microfluidic chip and three-dimensional entity technology, which is applied in drug screening, biochemical equipment and methods, compound screening, etc., can solve the problems of accuracy and reliability of analysis results, simple models, and poor integration of drug screening, etc., to achieve easy The effect of mass production, high-throughput detection, and miniaturization
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Example Embodiment
[0074] Embodiment 1 The structure of the chip of the present invention
[0075] The chip of the present invention is composed of a PDMS substrate and a cover sheet which are bonded together. Its structure is figure 1 As shown, the structure of the cover sheet is three parallel channels, and the parallel channels are connected by connecting channels. Among them, there are microchannels formed by drug perfusion and monolayer blood vessels (1), connecting channels of extracellular matrix (2), and microchannels with 14 "U"-shaped groove structures formed by three-dimensional tumor multicellular spheroids (3) ; The substrate has a shallower parallel channel that facilitates cell capture. The micro-tissue formed by the invention is very similar to the solid tumor micro-environment in the body, and the penetration of the drug in the micro-tissue can be observed in real time. Toxicity study. Cells that have undergone drug toxicity studies can flow out of the microchannels for subsequ
Example Embodiment
[0076] Example 2 Production of a chip of the present invention with PDMS as a substrate and a cover sheet
[0077] (1) Put the silicon wafer into concentrated H 2 SO 4 / H 2 O 2 (3 / 1) The solution was slightly boiled for 30 minutes, taken out after cooling, rinsed with deionized water until neutral, sonicated with ethanol for 5 minutes, and heated on a heating plate for 30 minutes.
[0078] (2) A thin layer of SU-8 photoresist is applied on the silicon wafer, and the thickness of the adhesive layer is about 50 μm by controlling the speed of the glue rejection. will have figure 2 The mask of the structure is attached to the silicon wafer, and ultraviolet light passes through the mask to expose the photoresist. Another spin on this silicon wafer with a thickness of about 50 μm will have image 3 The mask of the structure is attached to the silicon wafer, and is subjected to secondary exposure with ultraviolet rays, and the unexposed part is dissolved with a developing sol
Example Embodiment
[0081] Example 3 Using the chip prepared in Example 2 to form three-dimensional solid tumor microtissues
[0082] (1) If figure 1 , the channel (2) of the chip obtained by the present invention is perfused with basement membrane extract (BME) gel to simulate extracellular matrix, and then the channels (1) and (3) are modified with 1% (w / v) polyvinyl alcohol (PVA). ), preventing cells from sticking to PDMS.
[0083] (2) Set the concentration to 1×10 5 tumor cells / mL are perfused into the channel (3), so that the cells are uniformly and effectively captured in the "U"-shaped groove, and uniform tumor multicellular microspheres are formed after culturing for 48 hours.
[0084] (3) Set the concentration to 5×10 4 1 / mL human umbilical vein endothelial cells (HUVECs) flow into channel (1), place the chip vertically to make the cells grow on the BME glue, and place the chip horizontally after 2 hours to remove excess cells in the channel. Since BME has adhesion factors and growt
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