Tumor-targeted delivery carrier based on cell-derived micro-vacuoles, preparation method and application
A technology for tumor targeting and delivery vectors, which is applied in the field of preparation of tumor targeting delivery vectors and can solve the problem of low yield
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Embodiment 1
[0089] A method for preparing a tumor-targeted delivery carrier based on cell-derived microvesicles, the steps of which are:
[0090] 1. Conditioned medium: basal medium (HyClone, Waltham, MA, USA) was supplemented with 10% (v / v) fetal bovine serum (fetal bovine serum, FBS) (HyClone, Waltham, MA, USA) and 1% ( v / v) Antibiotics, add DSPE-PEG-Biotin (50ug / mL) and DSPE-PEG-Folate (5ug / mL) at the same time, store at 4°C to obtain conditioned medium.
[0091] 2. Use the conditioned medium obtained in step 1 for cell culture, culture under standard conditions for about 48 hours, replace the medium when the cells grow to a confluence of about 80%, and continue to culture with serum-free basal medium Cells, to promote the release of microvesicles from the cells, collect the cell culture supernatant after 46-50 hours of culture, and use for subsequent isolation to obtain functionalized microvesicles.
[0092] 3. Centrifuge the culture supernatant obtained in step 2 at 2000g for 20min ...
Embodiment 2
[0096] In vivo and in vitro biosafety testing of a tumor-targeted delivery carrier based on cell-derived microvesicles, the steps of which are as follows:
[0097] 1. Conditioned medium: basal medium (HyClone, Waltham, MA, USA) was supplemented with 10% (v / v) fetal bovine serum (fetal bovine serum, FBS) (HyClone, Waltham, MA, USA) and 1% ( v / v) Antibiotics, add DSPE-PEG-Biotin (50ug / mL) and DSPE-PEG-Folate (5ug / mL) at the same time, store at 4°C to obtain conditioned medium.
[0098] 2. Use the conditioned medium obtained in step 1 for cell culture, culture under standard conditions for about 48 hours, replace the medium when the cells grow to a confluence of about 80%, and continue to culture with serum-free basal medium Cells, to promote the release of microvesicles from the cells, collect the cell culture supernatant after 46-50 hours of culture, and use for subsequent isolation to obtain functionalized microvesicles.
[0099] 3. Centrifuge the collected supernatant obtain...
Embodiment 3
[0105]A tumor-targeted detection in vivo and in vitro of a cell-derived microvesicle-based tumor-targeted delivery carrier, the steps of which are as follows:
[0106] 1. Conditioned medium: basal medium (HyClone, Waltham, MA, USA) was supplemented with 10% (v / v) fetal bovine serum (fetal bovine serum, FBS) (HyClone, Waltham, MA, USA) and 1% ( v / v) Antibiotics, DSPE-PEG-Biotin (50 μg / mL) and DSPE-PEG-Folate (5 μg / mL) were added at the same time, stored at 4° C. to obtain conditioned medium.
[0107] 2. Use the conditioned medium obtained in step 1 for cell culture, culture under standard conditions for about 48 hours, replace the medium when the cells grow to a confluence of about 80%, and continue to culture with serum-free basal medium Cells, to promote the release of microvesicles from the cells, collect the cell culture supernatant after 46-50 hours of culture, and use for subsequent isolation to obtain functionalized microvesicles.
[0108] 3. Centrifuge the collected su...
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