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Culture method of exosome with high immunoregulation capability

A culture method and technology of regulation ability, applied in the field of biomedicine, can solve the problems of different immune regulation ability of exosomes and large difference in the quality of mesenchymal stem cells, and achieve the effect of enhancing immune regulation ability

Pending Publication Date: 2022-02-18
山东佰傲干细胞生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the large differences in the quality of mesenchymal stem cells from different tissue sources, different individual sources, and different batches, the immunomodulatory capabilities of the obtained exosomes are different.

Method used

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  • Culture method of exosome with high immunoregulation capability
  • Culture method of exosome with high immunoregulation capability

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Culture method of exosomes with high immune regulation ability

[0024] 1. Isolation and culture of mesenchymal stem cells

[0025] Take the umbilical cord tissue out of the collection bottle, wash it twice in alcohol, transfer it to sterile PBS to remove connective tissue, congestion, etc., and then wash it thoroughly to remove blood stains. Transfer the umbilical cord into a large petri dish, cut it into 3-4cm lengths and then separate it longitudinally, so that the umbilical cord tissue becomes tubular and flake-like. The umbilical cord tissue is further separated and cut into 2-3mm pieces 3 Tissue pieces of different sizes were planted one by one in a petri dish pre-coated with α-MEM medium containing 10% fetal bovine serum, and placed at 37 °C with a volume fraction of 5% CO 2 In a saturated humidity incubator, add 0.5 mL of medium every 24 hours, change the medium after 72 hours, and change the medium twice a week. When the cells grow to 80-85% conflu...

Embodiment 2

[0031] Example 2: Effects of exosomes on Treg cells after CD4+ T cells were treated

[0032] Peripheral blood lymphocytes were separated, and CD4+ T cells were sorted by magnetic beads. Divided into three groups, the blank control group was a single T cell, the control group co-cultured exosomes derived from mesenchymal stem cells without the combination of protein factors and T cells, and the experimental group used the exosomes in Example 1 and T cells co-cultivate. After 72 hours, the number of cells lost was used to detect the percentage of Treg cells.

[0033] figure 2 It is the expression of Treg cells. It can be seen from the figure that the exosomes of the experimental group and the control group can promote the transformation of T cells into Treg cells, but the promotion effect of the experimental group is significantly increased.

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Abstract

The invention provides a culture method of an exosome with high immunoregulation ability, which comprises the following steps: pretreating mesenchymal stem cells by using a prostaglandin E2, tumor necrosis factor-alpha and interferon-gamma protein factor combination to enhance the immunoregulation ability of the mesenchymal stem cells, and then extracting the exosome to further improve the immunoregulation capability of the exosome.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a method for cultivating exosomes with high immune regulation ability. Background technique [0002] Exosomes are a kind of vesicles secreted by cells to the outside of the cell. Its size is 30-150nm. Molecular transfer between. Exosomes are widely present in cell culture supernatants and various body fluids, including blood, lymph, saliva, urine, semen, milk, etc., and also in tissue samples, such as brain tissue, muscle tissue, adipose tissue, etc. . Exosomes can directly activate recipient cells through cell membrane receptors, and can also transport proteins, mRNA, miRNA, lncRNA, and even organelles into recipient cells to participate in intercellular communication. [0003] The ability of exosomes to regulate immunity is mainly due to the fact that exosomes are rich in numerous biologically active molecules, such as chemokines, inflammatory factors, signal...

Claims

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Application Information

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IPC IPC(8): C12N5/0775A61K35/28A61P37/04
CPCC12N5/0663C12N5/0667C12N5/0668C12N5/0665A61K35/28A61P37/04C12N2501/02C12N2501/25C12N2501/24C12N2500/90
Inventor 高迎凤程蕊苹蒲锋星
Owner 山东佰傲干细胞生物技术有限公司