Method for increasing yield of mesenchymal stem cell exosomes, and application of method

A technology for stem cells and exosomes, applied in the field of increasing the production of exosomes in mesenchymal stem cells, can solve the problem of low efficiency of exosome isolation and purification, increase of exosome production cost, and low exosome yield hindering clinical application. and other problems to achieve the effect of reducing culture cost, saving process cost and time, and improving production

Inactive Publication Date: 2021-02-09
陈传果
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, exosomes are mostly obtained by planar culture of mesenchymal stem cells, and the low yield of exosomes hinders their clinical application.
At the same time, the separation and purification efficiency of exosomes is low, which also increases the production cost of exosomes.

Method used

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  • Method for increasing yield of mesenchymal stem cell exosomes, and application of method
  • Method for increasing yield of mesenchymal stem cell exosomes, and application of method
  • Method for increasing yield of mesenchymal stem cell exosomes, and application of method

Examples

Experimental program
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Effect test

Embodiment 1

[0028] Embodiment 1 A method for increasing the production of exosomes from mesenchymal stem cells, comprising the following steps:

[0029] 1. The materials, reagents, instruments, etc. used in the examples can be obtained from commercial sources unless otherwise specified.

[0030] 2. Isolation and Identification of Umbilical Cord Mesenchymal Stem Cells

[0031] 1) Obtain a 30 cm umbilical cord from a healthy puerpera without knowing it. After collection in the operating room, soak it in sterile PBS and transport it to the laboratory. Soak it in 75% alcohol for 1 minute and rinse it, then put it on a 10 cm plate, rinse it with sterile PBS 3 times.

[0032] 2) Cut the umbilical cord into 5 cm pieces, tear the umbilical cord with tweezers and carefully remove the blood vessels (including 2 arteries and 1 vein), and carefully tear the umbilical cord into filaments.

[0033] 3) Transfer the filaments to a 50 mL centrifuge tube, add 10 mL PBS and mix well, then insert surgical ...

Embodiment 2

[0049] Example 2 Clinical application of a method for increasing the production of exosomes from mesenchymal stem cells

[0050] The isolated and purified exosomes were counted according to 10 7 Intra-articular injection was performed on 10 patients with osteoarthritis at a dose per kg, and the curative effect was evaluated at 1, 2, and 3 months after treatment. The results showed that none of the 10 patients had adverse reactions, the knee pain was relieved, the range of motion of the joints was increased, the thickness of the articular cartilage was increased, and the quality of life was improved.

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Abstract

The invention provides a method for increasing the yield of mesenchymal stem cell exosomes, and an application of the method. According to the method, a microcapsule is used as a growth system of mesenchymal stem cells, the mesenchymal stem cells are cultured under the three-dimensional condition, the culture environment is closer to the body, and therefore the yield of exosomes secreted by the mesenchymal stem cells can be increased conveniently. A semi-permeable membrane of the microcapsule can effectively intercept the exosomes, the concentration of the exosomes in the microcapsule is improved, and therefore the process cost and time are saved for downstream separation and purification; and exosomes derived from the serum of a culture medium can be prevented from entering the microcapsule, a specific commercial serum-free culture medium is prevented from being used, and therefore the culture cost is reduced. The method for increasing the yield of the mesenchymal stem cell exosomes through three-dimensional culture provided by the invention is expected to solve the technical bottleneck of insufficient exosome yield.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for increasing the exosome production of mesenchymal stem cells and its application. Background technique [0002] Exosomes are small vesicles with a diameter of 30-150 nm secreted by cells, which contain biomolecules such as cell-related proteins and nucleotides, and play an important role in cell communication. Recent studies have found that exosomes derived from stem cells can effectively transport biologically active substances such as mRNA, microRNA, and proteins, and have important biological functions such as reducing apoptosis, reducing inflammation, promoting angiogenesis, inhibiting fibrosis, and improving tissue repair potential. , there is a good clinical application prospect in regulating tissue regeneration. In a variety of disease / injury models, mesenchymal stem cell-derived exosomes have been shown to restore tissue function after intervention, and have simi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775C12N11/10C12N11/096C12N11/04A61K35/28A61P19/08A61P17/00
CPCC12N5/0665C12N5/0667C12N5/0663C12N11/10C12N11/08C12N11/04A61K35/28A61P19/08A61P17/00C12N2509/00
Inventor 陈传果
Owner 陈传果
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