Choroidal neovascularization targeting nanoparticle coated with macrophage membrane and preparation method of choroidal neovascularization targeting nanoparticle
A technology targeting nanoparticles and neovascularization, which is applied in the field of choroidal neovascularization targeting nanoparticles and its preparation, can solve the problems that there is no intravenous nano-preparation for the treatment of retinal diseases, poor drug compliance of patients, etc., and achieve high drug loading efficiency , strong drug-loading ability, and efficient treatment of diseases
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Embodiment 1
[0046] (1) Extraction of macrophages: 25 g of C57 male mice were intraperitoneally injected with 1 mL of 4% thioglycolate broth. After 3 days, the mice were sacrificed, the whole body was immersed in 75% ethanol for disinfection, and 5 mL of medium was injected twice to extract peritoneal macrophages. Cells were extracted at 37°C. 5%CO 2 Cultivate to 80%-90% under certain conditions;
[0047] (2) Collect the obtained cells into a centrifuge tube, centrifuge 3 times at 500g×5min, freeze and thaw repeatedly 3 times with liquid nitrogen to lyse the cells, and then repeatedly grind 20 times with a Dawns homogenizer to further break the cells;
[0048] Collect the cell lysate in a centrifuge tube, centrifuge at 3200g×5min at 4°C to remove large cell debris;
[0049] Collect the supernatant, centrifuge at 20000g×25min at 4°C to remove organelles;
[0050] Collect the supernatant, centrifuge at 100000g×60min, 4°C, discard the supernatant, and the resulting white precipitate is the...
Embodiment 2
[0059] (1) Extraction of macrophages: SD rats were injected intraperitoneally with 1 mL of 4% thioglycollate broth, sacrificed 3 days later, immersed in 75% ethanol for disinfection, and injected 5 mL of medium twice to extract peritoneal macrophages Cells were extracted at 37°C. 5%CO 2 Cultivate to 80%-90% under certain conditions;
[0060] (2) Collect the obtained cells into a centrifuge tube, centrifuge 3 times at 500g×5min, freeze and thaw repeatedly 3 times with liquid nitrogen to lyse the cells, and then repeatedly grind 20 times with a Dawns homogenizer to further break the cells;
[0061] Collect the cell lysate in a centrifuge tube, centrifuge at 3200g×5min at 4°C to remove large cell debris;
[0062] Collect the supernatant, centrifuge at 20000g×25min at 4°C to remove organelles;
[0063] Collect the supernatant, centrifuge at 100000g×60min, 4°C, discard the supernatant, and the resulting white precipitate is the cell membrane; resuspend the cell membrane pellet i...
Embodiment 3
[0069] Preparation of membrane-coated nanoparticle preparations of rapamycin-loaded macrophages labeled with cell membrane orange-red fluorescent probe DiI.
[0070] (1) Obtain macrophages directly from the mouse mononuclear macrophage leukemia cell line Raw264.7 cells, at 37°C, 5% CO 2Cultivate to 80%-90% under certain conditions;
[0071] (2) Scrape the macrophages obtained in (1), collect them in a centrifuge tube, centrifuge 3 times at 500g×5min, freeze and thaw them 3 times with liquid nitrogen to lyse the cells, and grind them repeatedly with a Dawns homogenizer 20 times to further break the cells; collect the cell lysate in a centrifuge tube, centrifuge at 3200g×5min, 4°C to remove large cell debris;
[0072] Collect the supernatant, centrifuge at 20000g×25min at 4°C to remove organelles;
[0073] Collect the supernatant, centrifuge at 100000g×60min, 4°C, discard the supernatant, and the resulting white precipitate is the cell membrane; resuspend the cell membrane pel...
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