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Method for strengthening immune-suppression function of mesenchymal stem cells

A technology of mesenchymal stem cells and immunosuppression, applied in the direction of adding compounds to stimulate growth, etc., to achieve the effect of small side effects, high efficiency, and low cost

Active Publication Date: 2013-02-20
SHANDONG QILU STEM CELL ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In order to solve the above problems that mesenchymal stem cells have low immunosuppressive ability and require a large amount of cells to function, the present invention provides a A method for enhancing the immunosuppressive function of mesenchymal stem cells by up-regulating key proteins in the regulation process

Method used

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  • Method for strengthening immune-suppression function of mesenchymal stem cells
  • Method for strengthening immune-suppression function of mesenchymal stem cells
  • Method for strengthening immune-suppression function of mesenchymal stem cells

Examples

Experimental program
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Effect test

Embodiment 1

[0037] 1. Enhance the immunosuppressive function of MSC

[0038] Extraction of umbilical cord mesenchymal stem cells

[0039] Neonatal umbilical cords authorized with maternal consent were used as a source of mesenchymal stem cells. The neonatal umbilical cord was washed three times in normal saline containing 1% double antibody to remove the blood stains on the surface, and then cut into small pieces about 1 cm long. Use ophthalmic scissors to cut the umbilical cord longitudinally along the direction parallel to the blood vessels, and peel off the 2 umbilical arteries and 1 umbilical vein from the umbilical cord. The remaining part of Huatong glue was fully washed 3 times with normal saline containing 1% double antibody, and cut into pieces to about 1mm 3 size.

[0040] Evenly spread the shredded tissue blocks on the 75cm 2 Add 5mL culture medium to the culture bottle, place it at 37℃, 5% CO 2 Cultivate in an incubator for 5-10 days. The medium composition is: 10% (volu...

Embodiment 2

[0056] On the basis of Example 1, change the concentration of the three culture factors in the medium to IFN-γ (Sigma) 1ng / mL, IL-1β (Sigma) 1ng / mL, progesterone (Sigma) 1μg / mL, other operations Completely consistent with embodiment 1. The effect of mixed lymphocyte reaction between MSC activated by cytokines and peripheral blood lymphocytes is as follows: Figure 4 , the inhibition rate of the obtained MSCs on lymphocyte proliferation was increased by 30%. Quantitative PCR was used to determine the expression effect of immunosuppressive proteins such as Figure 5 , the expression of immunosuppressive proteins in MSC cells was increased by 2%-200%. The obtained MSCs were assayed with an enzyme-linked immunosorbent assay kit for the secretion effects of anti-inflammatory factors and pro-inflammatory factors as follows: Figure 6 , the ability of cells to secrete anti-inflammatory factors increased by 10%-50%, and the ability to secrete pro-inflammatory factors did not change...

Embodiment 3

[0058] On the basis of Example 1, change the concentration of the three culture factors in the medium to IFN-γ (Sigma) 600ng / mL, IL-1β (Sigma) 200ng / mL, progesterone (Sigma) 100μg / mL, other operations Completely consistent with embodiment 1. The effect of mixed lymphocyte reaction between MSC activated by cytokines and peripheral blood lymphocytes is as follows: Figure 7 , the inhibition rate of the obtained MSCs on lymphocyte proliferation was increased by 25%. Quantitative PCR was used to determine the expression effect of immunosuppressive proteins such as Figure 8 , the expression of immunosuppressive proteins in MSC cells was increased by 5%-80%. The obtained MSCs were assayed with an enzyme-linked immunosorbent assay kit for the secretion effects of anti-inflammatory factors and pro-inflammatory factors as follows: Figure 9 , the ability of cells to secrete anti-inflammatory factors increased by 15%-50%, and the ability to secrete pro-inflammatory factors slightly ...

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Abstract

The invention relates to the technical field of mesenchymal stem cells for cell therapy, in particular to a method for strengthening an immune-suppression function of the mesenchymal stem cells. Separated mesenchymal stem cells are sub-cultured and then cultured in a culture medium containing IFN-gamma, IL-1beta and progesterone. After subjected to co-cultivation and activation of the IFN-gamma, theIL-1beta and the progesterone, the suppression ratio of the MSC cells to lymphocyte proliferation is improved by 2-7 times, the expression of immune-suppression protein in the MSC cells is improved by 2-12 times, the capacity of cell secretion anti-inflammatory cytokines is improved by 5-20 times, and the capacity of secretion proinflammatory cytokines is reduced by 10%-30%, and the immune-suppression activity of the MSC cells is increased greatly.

Description

technical field [0001] The invention relates to the technical field of mesenchymal stem cells in the field of cell therapy, and in particular to a method for enhancing the immunosuppressive function of mesenchymal stem cells. Background technique [0002] Mesenchymal stem cells (MSCs) are a type of adult stem cells that are derived from mesoderm, capable of self-renewal, and have the potential to differentiate into osteoblasts, cartilage cells, and adipocytes. Because MSCs are easy to isolate and culture, have strong proliferative ability, low immunogenicity, and there are no ethical problems in the wide range of sources, this makes their application in cell therapy have advantages that other stem cells do not have. MSC not only has low immunogenicity, but also has the function of regulating immune function. MSC can not only inhibit the immune response by inhibiting the maturation of dendritic cells and inhibiting the immune function of T lymphocytes, B lymphocytes and NK...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/38
Inventor 刘小盾李栋
Owner SHANDONG QILU STEM CELL ENG
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