Nanofiber membrane for capturing folate receptor high-expression cancer cells and preparation method and application thereof
A nanofiber membrane and folic acid receptor technology, applied in fiber types, fiber treatment, laboratory containers, etc., to achieve simple and quick modification, high capture rate, and simple production
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Embodiment 1
[0050] Weigh 2.0g of PLGA, dissolve it in a mixed solvent of THF and DMF (the volume ratio of THF (tetrahydrofuran) and DMF (N-N-dimethylformamide) is 3:1), under magnetic stirring, until PLGA is completely dissolved, and the concentration of spinning solution obtained is 25% (1g / 4mL). Then draw the spinning solution into the syringe, and use a high-voltage electrospinning machine to perform electrospinning. The spinning conditions are: temperature 25°C, humidity 40-50%, voltage 20Kv; receiving distance 15cm; liquid flow rate 0.3mL / h. Wherein the receiving plate is a clean glass slide or a circular cover glass adhered to the glass slide. Dry in a fume hood for 12-24 hours. Scanning Eelectron Microscope (SEM) results showed that the obtained nanofibers had a smooth surface and a uniform diameter, with an average fiber diameter of 850.2 nm ( figure 2 ).
[0051] Then modify FA to the surface of PEI, dissolve 4.41mg of FA in 5mL of dimethyl sulfoxide DMSO, use ultrasound for ...
Embodiment 2
[0054] In order to verify the specific capture effect of the fiber material, human oral epithelial cells (KB cells) with high expression of folate receptors were used as model cells to test the effect of the prepared FA-modified functionalized nanofibers on specific capture of KB cells.
[0055] The obtained FA-modified circular coverslip covered with nanofibers was removed from the glass slide, and then the samples were placed into a 12-well culture plate. Samples at different time points were placed in different culture plates, and each sample had 3 parallel samples. After 1 hour of ultraviolet irradiation, 400 μL of serum-free medium was added to the well plate to soak the fiber slides for 30 minutes. Then the culture medium was aspirated, and the prepared serum-free cell suspension was added to the well plate, the cell concentration was 250 cells / mL, and the suspension volume was 400 μL to ensure that 100 cells were added to each well plate. Then let it stand for 10, 20, ...
Embodiment 3
[0057] Prepare polydimethylsiloxane (PDMS) chips (e.g. Image 6 Shown): Mix the prepolymer and curing agent evenly at a mass ratio of 10:1, remove air bubbles in a vacuum, pour the PDMS solution into a petri dish with a silicon wafer mold, and cure in an oven at 60°C. The folic acid-modified PLGA nanofiber membrane is cut into a size similar to the cell capture channel of the chip, and then the cut nanofiber membrane slide is subjected to ion treatment together with the microfluidic chip. Among them, the glass slide covered with nanofiber membrane is only treated on the glass surface, and the fiber membrane is temporarily shielded by a slightly larger PDMS membrane to prevent plasma damage to the fiber membrane. Immediately after treatment, the two are bonded under a certain pressure. Wrap the bonded device in a sealing film to prevent contamination and store it away from light. Before the cell capture experiment, use epoxy resin AB glue to glue the medical infusion tube at ...
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Abstract
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