Hybrid mesenchymal stem cell exosome drug delivery system as well as preparation method and application thereof
A mesenchymal stem cell and delivery system technology, applied in the field of hybrid mesenchymal stem cell exosome drug delivery system and its preparation, can solve the problems of tumor drug resistance, poor prognosis of tumor treatment, etc. To improve the efficiency and improve the effect of tumor targeting inhibition
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[0057] A method for preparing a hybrid mesenchymal stem cell exosome drug delivery system as described above, the steps are as follows:
[0058] S1 Dissolve cholesterol components and phospholipid materials in an organic solvent at a weight ratio of 5-20:80-95, then add fat-soluble antineoplastic drugs, and bathe in a water bath of 35-55°C to dissolve the cholesterol components and phospholipid materials evenly;
[0059] Under the condition of S2 water bath, rotate and vacuum to remove the organic solvent, so that a honeycomb film is formed on the inner wall of the container, and place it in a vacuum drying oven for 8-12h;
[0060] S3 Take the film obtained in step S2, add a pH buffer solution in which a water-soluble antitumor drug is dissolved, and rotate in a water bath at 40-60° C. to completely dissolve the film;
[0061] S4 Ultrasound or homogenization to prepare drug-loaded liposomes with a median diameter of 60-200 nm;
[0062] S5 uses the preparation technology of pH...
Embodiment 1
[0074] Example 1 Preparation of tirapazamine-hydralazine dual-loaded hybrid mesenchymal stem cell exosomes
[0075] (1) Preparation of exosome stock solution
[0076] Take 1ml of fresh animal mesenchymal stem cell sample into a cell culture bottle, add DMEM / F12 medium containing 10% fetal bovine serum, and culture in a constant temperature incubator at 37°C and 5% CO2; respectively, at 24, 48 and 72 hours Change the medium of the culture bottle completely, and discard non-adherent cells such as red blood cells and adipocytes; change the medium every three days, perform primary culture for 10-15 days, and conduct the first passage, and observe the growth of adherent mesenchymal stem cells after 4-7 days When reaching 80-90%, subculture again; get the bone marrow mesenchymal stem cells of the 3rd-6th generation ( figure 1 ), collected cell supernatant 100ml, divided into centrifuge tubes, centrifuged at 300×g for 10min, took the supernatant, centrifuged at 2000×g for 10min, too...
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