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Recombinant mesenchymal stem cells, preparation method and application thereof

A mesenchymal stem cell and cell culture technology, applied in the field of recombinant mesenchymal stem cells and their preparation, can solve the problems of low ability and low efficiency of secreting anti-inflammatory molecules

Inactive Publication Date: 2016-06-15
SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In summary, traditional mesenchymal stem cells have low homing efficiency to disease or inflammatory injury sites, and low ability to secrete anti-inflammatory molecules

Method used

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  • Recombinant mesenchymal stem cells, preparation method and application thereof
  • Recombinant mesenchymal stem cells, preparation method and application thereof
  • Recombinant mesenchymal stem cells, preparation method and application thereof

Examples

Experimental program
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Effect test

preparation example Construction

[0070] The preparation method of the above-mentioned recombinant mesenchymal stem cells is simple and efficient. Using the mRNA transfection method, PSGL-1 and SLeX can improve the ability of mesenchymal stem cells to target and homing cells in transplantation therapy, and the recombined mesenchymal stem cells It can secrete IL-10 molecules with high-efficiency anti-inflammatory effect at the site of inflammation and improve the therapeutic effect.

[0071] In addition, a drug targeting carrier is also provided, and the targeting carrier includes the recombinant mesenchymal stem cells described in any one of the above.

[0072] When the recombinant mesenchymal stem cells are used as drugs or targeted carriers for treating multiple sclerosis and other diseases, the therapeutic effect can be increased, and a new and highly effective therapeutic drug is provided for the treatment of immune diseases.

Embodiment 1

[0075] Example 1 Preparation of Human Bone Marrow Recombinant Mesenchymal Stem Cells

[0076] (1) The first mRNA, the second mRNA and the third mRNA are respectively synthesized by genetic engineering to obtain the first mRNA for expressing P-selectin glycoprotein ligand-1 and the first mRNA for expressing sialylated Lewis oligosaccharide-X A second mRNA for the antigen and a third mRNA for expressing said interleukin-10.

[0077] (2) To prepare human bone marrow mesenchymal stem cells, replace with fresh preheated medium (DMEM) 1 hour before gene transfection.

[0078] (3) Prepare serum-reduced medium (Opti-MEM, purchased from GibcoLifeTechnology) and RNAiMAX transfection reagent (purchased from Lipofectamine), and dissolve the first messenger RNA, second messenger RNA, and third messenger RNA in the reduced-serum medium , and then add RNAiMAX transfection reagent, mix well and let stand for 15 minutes to obtain transfection mixture. The amount of serum-reduced medium was c...

Embodiment 2

[0080] Example 2 Preparation of recombinant human umbilical cord mesenchymal stem cells

[0081] The transfected cells were human umbilical cord mesenchymal stem cells, and the rest of the conditions were the same as in Example 1 to obtain recombinant human umbilical cord mesenchymal stem cells expressing PSGL-1, SLeX and IL-10.

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Abstract

The invention discloses recombinant mesenchymal stem cells, a preparation method and application thereof. A first messenger RNA (ribonucleic acid) for expressing P lectin glycoprotein ligand-1, a second messenger RNA for expressing sialated Lewis oligosaccharide-X antigen and a third messenger RNA for expressing interleukin-10 are transfected into mesenchymal stem cells to obtain the recombinant mesenchymal stem cells.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a recombinant mesenchymal stem cell and its preparation method and application. Background technique [0002] Immune diseases (immune diseases) refer to the diseases caused by the imbalance of immune regulation and affecting the immune response of the body. Immunological diseases in a broad sense include structural or functional abnormalities of the immune system caused by congenital or acquired causes. Among them, multiple sclerosis (MS) is a common inflammatory demyelinating disease of the central nervous system mediated by autoimmune reactions. The lesions are mostly found in the white matter of the brain and spinal cord, showing multifocal inflammatory cell infiltration, demyelination, and neuronal damage, which can lead to sensory, motor, consciousness, and neurocognitive dysfunction. The disease mostly occurs in young adults aged 20 to 40, with a high recurrence rate and disa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10A61K48/00A61K47/48A61K38/20A61K35/28A61P37/02A61P3/00A61P29/00
CPCA61K35/28A61K38/20A61K48/00C12N5/10C07K14/435A61K38/2066A61K48/005C07K14/5428
Inventor 曾宪卓
Owner SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD
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