Stem cells with enhanced immunoregulation capability, preparation method thereof, composition and application thereof

An immune regulation and stem cell technology, applied in the field of biomedicine, can solve problems such as ineffective treatment of autoimmune diseases

Active Publication Date: 2019-12-13
CAPITAL UNIVERSITY OF MEDICAL SCIENCES +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, drugs that target and inhibit T cells and B cells cannot effectively treat autoimmune diseases, but autologous or allogeneic MSCs tra

Method used

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  • Stem cells with enhanced immunoregulation capability, preparation method thereof, composition and application thereof
  • Stem cells with enhanced immunoregulation capability, preparation method thereof, composition and application thereof
  • Stem cells with enhanced immunoregulation capability, preparation method thereof, composition and application thereof

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preparation example Construction

[0047] [Preparation method of stem cells with enhanced immune regulation ability]

[0048] The second aspect of the present invention provides a method for preparing stem cells with enhanced immune regulation ability (sometimes referred to as "preparation method of the present invention" or "method of the present invention" for short in the present invention), including using, for example, genetic engineering to reduce The expression level of the Id3 gene in the cell or the step of reducing the activity of the ID3 protein.

[0049] In some embodiments, the preparation method of the present invention includes the step of reducing the expression level of the Id3 gene in the cells by using RNA interference technology. RNA interference technology is a technology in which exogenous and endogenous double-stranded RNA induces RNA-specific degradation of homologous target genes in vivo, resulting in post-transcriptional gene silencing. A technique using siRNA or shRNA is preferred. ...

Embodiment

[0084] 1. BMMSCs derived from human stem cell bank were modified by Id3

[0085] Preparation of Id3 siRNA knockout BMMSCs

[0086] 1) Synthesize Id3 siRNA, the sequence is GGAGCTTTTGCCACTGACT, or GCTTGCTGGACGACATGAA, or GCCAGGTGGAAATCCTACA

[0087] 2) Shop 5×10 5 Put BMMSCs cells per well into a 24-well plate, with 150 μL medium in each well, which is α-MEM complete medium;

[0088] 3) On the second day, 50 μL / well of OPTI-MEM was mixed with 1 μL Lipofectamin 2000, and incubated for 5 minutes;

[0089] 4) Mix 4 μL of synthetic si-Id3, siRNA control with another 50 μL / well of OPTI-MEM;

[0090] 5) Mix the above two mixtures and incubate at room temperature in the dark for 20 minutes;

[0091] 6) Suck off the original complete medium, and slowly add the mixture to the cell culture plate;

[0092] 7) After 6 hours of transfection, suck off the mixed solution, and replace with preheated complete culture medium without antibiotics. Three days later, collect the cells, extract ...

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Abstract

The invention discloses stem cells with enhanced immunoregulation capability, a preparation method thereof, a composition containing the same, and an application thereof. The immunomodulatory abilityof stem cells is enhanced by reducing the expression level of an Id3 gene in the stem cells or reducing the activity of ID3. The stem cells with enhanced immunoregulation ability are used for treatment of autoimmune-related diseases or conditions.

Description

technical field [0001] The present invention generally relates to the field of biomedicine, and specifically relates to a stem cell with immune regulation ability, a preparation method thereof, a composition containing the same, and a method for enhancing and detecting the immune regulation ability of the stem cell. Background technique [0002] Autoimmune diseases (autoimmune diseases, AD) is an immune response disorder, the body's immune system attacks self-antigens, resulting in tissue and organ damage and dysfunction, with high disability and mortality rates. Most ADs are multiple sclerosis, systemic sclerosis, systemic lupus erythematosus, rheumatoid arthritis, juvenile idiopathic arthritis, and immune cytopenias. The occurrence of autoimmune diseases is an extremely complex regulatory network. In addition to lymphocytes, such as T cells, B cells, dendritic cells and NK cells, participating in the pathogenesis of autoimmune diseases, mesenchymal stem cells (MSCs) Disor...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12N15/85G01N33/569
CPCC12N5/0662C12N5/0647C12N15/85G01N33/56966C12N2510/00C12N2810/85
Inventor 王松灵胡磊徐骏疾陈万军吴婷婷李艳
Owner CAPITAL UNIVERSITY OF MEDICAL SCIENCES
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