Mesenchymal stem cell for treating autoimmune diseases and preparation method and application of mesenchymal stem cell

A technology for autoimmune diseases and mesenchymal stem cells, applied in mesenchymal stem cells and its preparation method and application field

Active Publication Date: 2019-04-16
北京贝来生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, during the use of the above-mentioned antibodies, antibodies against monoclonal antibodies were detected in the serum of som...

Method used

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  • Mesenchymal stem cell for treating autoimmune diseases and preparation method and application of mesenchymal stem cell
  • Mesenchymal stem cell for treating autoimmune diseases and preparation method and application of mesenchymal stem cell
  • Mesenchymal stem cell for treating autoimmune diseases and preparation method and application of mesenchymal stem cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Embodiment 1: IL17RA-ECD: Fc fusion protein gene vector construction

[0065] We designed IL17RA-ECD and IL17RA-ECD:Fc gene plasmids through Genebank, and constructed IL17RA-ECD and IL17RA-ECD:Fc fusion protein lentiviral expression plasmids using the fourth-generation lentiviral vector system (LV-IL17RA and LV-IL17RA:Fc ), and LV-EGFP was used as a control virus (LV-null). ( figure 1 Lentiviral expression plasmid constructs for LV-EGFP, LV-IL17RA and LV-IL17RA-Fc).

Embodiment 2

[0066] Example 2: Establishment of human umbilical cord mesenchymal stem cell seed bank

[0067] Human umbilical cords were obtained from healthy full-term pregnancy cesarean section pregnant woman donors, provided by qualified national second-level hospitals, and pregnant women or their family members signed informed consent and umbilical cord collection registration forms. The extraction, cultivation and establishment of cell bank of umbilical cord mesenchymal stem cells are all completed in our company's GMP production workshop. After the cells are separated, cultured, subcultured and expanded to the second generation (P2), the exogenous microorganisms, viruses, endotoxins, etc. are detected; at the same time, the immune phenotype, differentiation ability and cell biological efficacy of the cells are detected. Qualified cells were used as seed bank cells and stored in a liquid nitrogen tank at -196°C.

Embodiment 3

[0068] Example 3: IL17RA-ECD: Fc Gene Modified Mesenchymal Stem Cells

[0069] The above-mentioned LV-IL17RA and LV-IL17RA: Fc fusion protein lentiviral expression plasmids and LV-EGFP were mixed with lentiviral pGag / Pol, pRev, pVSV-G and other framework plasmids respectively, introduced into 293T cells through LTX liposomes, and packaged to obtain Mature lentiviruses were harvested and infected with MSCs. After 24 hours, puromycin was added to select successfully infected cells.

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Abstract

The invention discloses a mesenchymal stem cell for treating autoimmune diseases. The mesenchymal stem cell specifically expresses a human interleukin-17 receptor analogue through a genetic modification method, thereby competitively inhibiting the combination of IL-17 and an IL-17 receptor. Therefore the mesenchymal stem cell can be used for inhibiting inflammation-related signaling pathways mediated by the IL-17 and inflammatory responses. The invention also discloses IL17RA fusion protein and a preparation method and application of the mesenchymal stem cell.

Description

technical field [0001] The invention relates to a mesenchymal stem cell for treating related autoimmune diseases by blocking the IL17 / IL17R conduction pathway and inhibiting the inflammatory response mediated by the IL17 pathway, as well as its preparation method and application, belonging to the field of biomedicine. Background technique [0002] Human interleukin 17 (hIL-17) is a pro-inflammatory cytokine produced by the activation of T helper cell TH17, which can recognize and mediate autoimmunity by directly or indirectly inducing a variety of cytokines, chemokines, inflammatory factors and antimicrobial proteins It plays an important role in inflammatory response, immune regulation, microbial and parasitic infection, and plays a key regulatory role in the human cytokine immune regulatory network. Six members of interleukin-17 (IL-17) have been discovered, namely: IL-17A, IL-17B, IL-17C, IL-17D, IL-17E (also known as IL-25) and IL-17 17F. The latest research shows that...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12N15/867C07K14/54A61K35/28A61P37/02A61P1/00A61P29/00A61P17/06A61P19/02A61P11/00
CPCA61K35/28A61P1/00A61P11/00A61P17/06A61P19/02A61P29/00A61P37/02C07K14/54C12N5/0662C12N15/86C12N2510/00C12N2740/15043
Inventor 刘广洋刘拥军李欣米一
Owner 北京贝来生物科技有限公司
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