A kind of brain targeting exosome, its preparation method and application
A technology of exosomes and brain, which is applied in the field of brain-targeted exosomes and its application in the treatment of brain diseases, can solve the problems of incomplete and single detection after traumatic brain injury, and achieve the goal of overcoming Difficulties in brain targeting, overcoming toxic side effects, overcoming ineffective effects
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[0029] Another aspect of the embodiments of the present invention also provides a method for preparing brain-targeted exosomes, including:
[0030] Make brain-targeted neural stem cells secrete exosomes with brain-targeted peptides;
[0031] collecting and purifying the exosomes;
[0032] The fluorescent material is allowed to enter the inner cavity of the exosome, thereby obtaining the brain-targeted exosome.
[0033] Among them, exosomes are derived from genetically engineered neural stem cells. The neural stem cells can be modified first by genetic engineering, so that the surface of the neural stem cell cell membrane carries a targeting peptide targeting the brain. Exosomes with brain-targeted peptides secreted by brain-targeted modified neural stem cells have the function of crossing the blood-brain barrier and can treat traumatic brain injury. Such a natural nanovesicle can achieve low Toxic side effects, efficient and precise treatment.
[0034] Fluorescent materials...
Embodiment
[0067] (1) Extraction of neural stem cells
[0068] Step 1: Take a Balb / C mouse about 4 weeks old, kill it by neck dislocation, cut off the head, take out the complete brain, and put it on the pre-cooled DMEM-F12 medium (containing 1% double antibody) on ice, Carefully peel off the hypothalamus (SVZ) area, put it on ice, peel off the meninges and blood vessels with forceps; chop the hypothalamus (SVZ) with a scalpel, put it into a 15 mL centrifuge tube containing 5 mL DMEM-F12 serum-free medium, add 0.5mL neural stem cells were digested with trypsin Accutase at 37°C for 20min and shaken every 5min. Centrifuge at 200g for 5 minutes to remove the supernatant; add 1mL DMEM-F12, culture medium to resuspend the cells, then add 20μl DNase, gently blow the tissue 10 times with a pipette gun, do not generate air bubbles during the blowing process. Add 1 mL of fresh medium and a small amount of DNase, pipette 15 times slowly on ice, let stand for 1-2min, pipette the supernatant into a...
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