A method for expanding neural stem cells through exosomes derived from endothelial cells

Abstract

The invention discloses a method for amplifying neural stem cells using exosomes derived from endothelial cells. After cultivating P3‑P4 generation human primary endothelial cells in a serum-free medium for 72 hours, the medium supernatant is collected; the cultured After the supernatant was centrifuged, it was filtered through a filter membrane, and polyethylene glycol 6000 with a final concentration of 12% was added and mixed; 12 hours after the mixing, it was centrifuged at a speed of 12000g / min for 1 hour to obtain exosomes; Add exosomes derived from human primary vascular endothelial cells to mouse neural stem cells, and incubate for 12 hours. After 5 days of culture, the self-renewal and proliferation capabilities of neural stem cells are enhanced; the method of the present invention is simple to operate, takes a short time, and can Maintain the normal stem cell characteristics and multi-directional differentiation potential of NSCs, and provide sufficient cell mass for stem cell replacement therapy using NSCs as the source of cells.

Description

Technical field: [0001] The invention belongs to the field of biomedicine, and relates to exosomes of endothelial cells and a method for amplifying neural stem cells, in particular to a method for exosomes derived from human primary umbilical vein endothelial cells in the expansion of neural stem cells in mice application. Background technique: [0002] After nearly a decade of research on vesicles, exosomes are a common means of communication between cells. Almost all cells in an organism can release exosomes, which are either taken up by surrounding cells, or circulate in various fluids, such as blood, urine, cerebrospinal fluid, and breast milk, and finally taken up by distant cells . Exosomes are derived from endosomes, which are lipid-encapsulated content carriers that can transport biologically active molecules such as proteins, nucleic acids, and lipids. Nucleic acids, proteins and lipids are packaged into exosomes and released after fusion with the cell membrane b...

AI Technical Summary

Problems solved by technology

All types of nerve cells are derived from neural stem cells (neural stem cells, NSCs). Brain injury will activate the proliferation and differentiation of NSCs to repair the damaged brain area, but this process is extremely inefficient. In addition, the number of NSCs in the brain is limited, so endogenous repair is far away. Inability to replenish large numbers of damaged nerve cells

Exogenous NSCs provide opportunities for the treatment of such diseases, but the current methods for expanding NSCs have obvious defects, such as the potential cancer risk of induced pluripotent stem cells, ethical issues in embryonic stem cell therapy, and cytokine expansion of NSCs cellular aging

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