Chemotherapy drug loaded and TIGIT overexpressed engineered drug-loaded cell membrane vesicle as well as preparation method and application thereof
A chemotherapeutic drug and cell membrane technology, applied in botany equipment and methods, biochemical equipment and methods, cells modified by introducing foreign genetic material, etc., can solve the problem of short circulation time in the body, easy to be degraded and cleared, unstable properties, etc. problems, to achieve the effect of improving bioavailability, effective enrichment, and high biosafety
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[0049] 1. Preparation of engineered drug-loaded cell membrane vesicles
[0050] 1. Packaging lentivirus. Culture HEK-293T cells (purchased) in a cell incubator at 37°C, add the mixed target plasmid (plasmid encoding TIGIT gene), packaging plasmid and lipo2000 (liposome) in a ratio of 1:1.2~1.5:2~4 2000). Change the medium after 5-9 hours, continue to culture with normal complete medium, and observe the transfected HEK-293T cells under a fluorescence microscope at different times. The result is as figure 1 As shown, it shows that HEK-293T cells are efficiently packaging and producing lentivirus.
[0051] The culture supernatant was collected and centrifuged to remove cells. Continue to collect the supernatant and filter through a 0.45 μm filter to remove cell debris in the supernatant. Then mix the filtrate with the virus purification reagent (Lenti Concentrator) at a volume ratio of 1:4, centrifuge at 3000g~4000g for 20-30min after incubation, remove the supernatant, and ...
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