Method for efficiently producing extracellular vesicles

Abstract

The invention discloses a method for efficiently producing extracellular vesicles. The method for producing the extracellular vesicles provided by the invention comprises the following steps: mixing cells with a biocompatible material to obtain a mixed solution; dispersing the mixed solution in a biological inert solution to form a biocompatible three-dimensional body; and culturing the biocompatible three-dimensional body in a cell culture medium, and collecting the supernatant to obtain the extracellular vesicles. According to the method, the cells are losslessly loaded to form a three-dimensional body, a good cell communication microenvironment is constructed, the efficiency of secreting extracellular vesicles by the cells is effectively improved, and the maximum utility utilization of the cells is achieved. The method is high in universality, simple in process, short in time consumption and high in uniformity, and can be used as a technical platform for large-scale production of the extracellular vesicles. Original forms, structures and functions of cells can be reserved, cell viability cannot be reduced, cell apoptosis is caused, or phenotypes of the cells cannot be changed, and therefore functions of diagnosis, treatment and the like of extracellular vesicles cannot be weakened.

Description

technical field [0001] The present application relates to the field of cell technology, in particular to a method for efficiently producing extracellular vesicles. Background technique [0002] With the increasing research on extracellular vesicles (including exosomes) at home and abroad, a variety of methods have been proposed to increase the production of extracellular vesicles, including the use of bioreactor systems to provide a larger surface area, which is convenient for cells to process. Amplification to increase the production of extracellular vesicles, but this method itself cannot improve the efficiency of cells secreting vesicles, and under large-scale production conditions, the homogeneity of culture conditions is difficult to monitor; it also includes changing cell culture conditions methods, such as increasing the production of extracellular vesicles by applying mechanical stimulation, changing the pH of the cell environment, inducing cell hypoxia, and increasi...

AI Technical Summary

Problems solved by technology

However, the method of suspension or hanging drop culture is time-consuming and has large heterogeneity between batches

The method of using microcarriers to form cell microspheres requires a complicated and cumbersome microcarrier preparation process, and the formation of cell microspheres requires complex devices and long-term stirring and mixing, and requires precise control of the concentration and stirring conditions of cells seeded on microcarriers , preventing the early formation of cell aggregates impairing cell growth and properties

In addition, different microcarrier surface substrates can cause changes in cell phenotypes, such as material stiffness, nano-topography, and local curvature that may affect cell proliferation, decreased protein expression levels, etc.

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