Brain-targeting exosome and preparing method and application thereof

An exosome and brain-targeting technology, which is applied in the direction of pharmaceutical formulations, medical preparations of non-active ingredients, cardiovascular system diseases, etc., can solve the problems of extraction waste and affecting curative effect, and achieve integrity and brain-targeting Ability to improve, enhance the effect of gathering

Inactive Publication Date: 2019-08-13
ZHEJIANG UNIV
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This will lead to the waste of difficult-to-extract exosomes in use, affecting the efficacy

Method used

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  • Brain-targeting exosome and preparing method and application thereof
  • Brain-targeting exosome and preparing method and application thereof
  • Brain-targeting exosome and preparing method and application thereof

Examples

Experimental program
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Embodiment Construction

[0040] 1. Extraction and separation of exosomes secreted by stem cells

[0041] Stem cells were resuspended in culture medium to prepare cell suspension (2×10 5 cell / mL), cultured at 37°C for 48h under normal conditions (oxygen concentration 20%, carbon dioxide concentration 5%).

[0042] The culture medium contained basal medium DMEM / F12 (Gibco, product number 11330032), 2% cell culture supplement B27 (Gibco, product number 17504-044), 1 μg / mL basic fibroblast growth factor (bFGF, PeproTech, product number 450- 33-50), 2 μg / mL epidermal growth factor (EGF, PeproTech, Cat. No. 315-09-100).

[0043] Collect the stem cell culture medium, centrifuge at low speed, centrifuge at 3000g for 30min, and take the supernatant; then centrifuge at 10000g for 30min, take the supernatant, discard cell debris and cell organelles and other precipitates. The centrifuged cell culture fluid was centrifuged and concentrated at a speed of 4000g with a 100KD ultrafiltration tube. The concentrated...

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Abstract

The invention discloses a brain-targeting exosome and a preparing method and application thereof. The preparing method comprises the following steps of 1, culturing stem cells under a low-oxygen condition, and conducting targeting modification on the stem cells, wherein according to the low-oxygen condition, the temperature is 30-40 DEG C, the concentration of oxygen is 0.5-5%, and the concentration of carbon dioxide is 1-10%; 2, transferring the stem cells after targeting modification under a normal culture condition for culture, collecting a stem cell culture liquid, and conducting extracting and separating from the stem cell culture liquid to obtain the brain-targeting exosome. The obtained brain-targeting exosome can be applied to preparing drugs for treating cerebral ischemic stroke.According to the preparing method, through modification on the stem cells, the exosome is indirectly modified, the expression of the exosome on the membrane surface CXCR4 is improved, and thus the brain targeting performance of the exosome is improved.

Description

technical field [0001] The invention relates to the technical field of biological carrier targeted drug delivery, in particular to a brain-targeted exosome and its preparation method and application. Background technique [0002] In recent years, with the changes in the global environment and the transformation of human life style, cardiovascular and cerebrovascular diseases have gradually occupied the first place in the cause of human death. Among them, there are about 3.4 million new cerebrovascular diseases in China every year, and about 1.6 million new deaths from cerebrovascular diseases. The morbidity and mortality are still increasing year by year. Ischemic stroke is characterized by high morbidity, high mortality, and high disability. The most common type of ischemic stroke is the ischemia-hypoxic necrosis of local brain tissue caused by stenosis or occlusion of blood supply arteries in the brain and intracranial blood circulation disorder, which eventually leads to...

Claims

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Application Information

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IPC IPC(8): A61K47/46A61P9/10
CPCA61K47/46A61P9/10
Inventor高建青蒋心驰
OwnerZHEJIANG UNIV