Exosome preparation prepared from umbilical cord mesenchymal stem cells, and preparation method of exosome preparation

Abstract

The invention relates to an exosome preparation prepared from umbilical cord blood bone marrow mesenchymal stem cells, and a preparation method of the exosome preparation. The exosome is obtained froma cell culture containing at least 10<4> cells. When the cell culture reaches fusion, washing is performed in PBS, a KO-DMEM culture medium is added and the cell culture is put back into an incubatorcontaining carbon dioxide for culture, the cell culture is cultured for 2-20 days, and the exosome is prepared through the ultracentrifugation technology, size exclusion filtration, chemical precipitation, discontinuous density gradient, immunoaffinity, ultrafiltration and / or high performance liquid chromatography, and preferably through PEG precipitation. The exosome prepared by the method is stable, high in purity, and comprehensive in positive markers, and can be used for activating proliferation and regeneration, promoting wound healing, and other medical clinical uses.

Description

technical field [0001] The present invention relates to cell biology, and in particular to a preparation method for producing stable exosomes. Background technique [0002] Exosomes are cell-derived vesicles (Hong, C.S., Muller, L., Boyiadzis, M. and Whiteside, T.L., 2014. Isolation and characterization of CD34+ blast-derived exosomes inacute myeloid leukemia. PloS one, 9(8 ), p.e103310). They are found in biological fluids such as urine, plasma, and ascites. Exosomes are produced by inward budding of endosomal multivesicular bodies. The molecules transported by exosomes include proteins / glycoproteins expressed on the cell membrane as well as molecules and soluble factors present in the parental cell cytosol. Exosomes usually have a diameter of 40–100 nm and contain proteins, lipids, RNA and microRNAs (Zhang, Z., Wang, C., Li, T., Liu, Z. and Li, L., 2014. Comparison of ultracentrifugation and density gradient separation methods for isolating Tca8113 human tongue cancer ...

AI Technical Summary

Problems solved by technology

Many of the current various technologies have their drawbacks and are not suitable for clinical applications, including ultracentrifugation (time-consuming, unstable yield, low purity and damage to vesicles), density gradient centrifugation (complex steps, very time-consuming ), magnetic bead immunoassay (low efficiency, exosome biological activity is easily affected by pH and salt concentration, which is not conducive to downstream applications) and precipitation method (not high purity, uneven particle size)

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