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Composition for improving, preventing or treating skin diseases comprising induced pluripotent stem cell-derived mesenchymal stem cell and exosome derived therefrom

A technology of pluripotent stem cells and skin diseases, applied in the field of compositions for improving, preventing or treating skin diseases comprising mesenchymal hepatocytes derived from induced pluripotent stem cells and exosomes derived therefrom

Pending Publication Date: 2020-11-10
THE ASAN FOUND +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is currently no known direct specific treatment for atopic dermatitis, so there is an urgent need to develop new treatments for atopic dermatitis

Method used

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  • Composition for improving, preventing or treating skin diseases comprising induced pluripotent stem cell-derived mesenchymal stem cell and exosome derived therefrom
  • Composition for improving, preventing or treating skin diseases comprising induced pluripotent stem cell-derived mesenchymal stem cell and exosome derived therefrom
  • Composition for improving, preventing or treating skin diseases comprising induced pluripotent stem cell-derived mesenchymal stem cell and exosome derived therefrom

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0106] Example 1: Preparation and analysis of mesenchymal stem cells derived from induced pluripotent stem cells

[0107] In a state that does not use supporting cells (cultivated auxiliary cells), it contains knock-out xeno-free serum replacement, glutamine, non-essential amino acids, β-mercaptoethanol, antibiotics, and basic ingredients. Induced pluripotent stem cells (fibroblasts and peripheral blood mononuclear cells cultured in Asan Hospital Stem Cell Center, Seoul) were cultured in DMEM / F-12 medium with bFGF (basic fibroblast growth factor). (Peripheral Blood Mononuclear Cell, PBMC, induced pluripotent stem cell line of mesenchymal stem cells) was attached to a culture dish (dish) coated with vitronectin in advance, and added with 10% fetal bovine serum ( v / v), 5ng / ml basic fibroblast growth factor, 0.1mM minimum essential medium non-essential amino acids (MEMNEAA, Minimum Essential Media Non-Essential Amino Acids), β-mercaptoethanol (β-mercaptoethanol) ( 1×), 100 unit / ...

Embodiment 2

[0118] Example 2: Isolation and confirmation of exosomes derived from mesenchymal cells differentiated from induced pluripotent stem cells

[0119] The iMSCs whose characteristics of mesenchymal stem cells were confirmed were additionally cultured in a medium supplemented with 10% exosome-depleted bovine serum (Exosome-depleted FBS). After culturing for 72 hours, the culture medium of recovered iMSCs was centrifuged at 300×g for 10 minutes to remove remaining cells and cell residues. The supernatant was filtered with a 0.22 μm filter, and then centrifuged at 10,000×g and 4° C. for 70 minutes using a high speed centrifuge. Use an ultracentrifuge (ultracentrifuge) at 100,000×g, 4°C to centrifuge the centrifuged supernatant for 90 minutes, remove the supernatant, and dilute the remaining supernatant with phosphate buffered saline (PBS, phosphate buffered saline). Exosomes are used.

[0120] The number and size distribution of exosomes isolated from MSCs and iMSCs were confirmed...

Embodiment 3

[0122] Example 3: Preparation and administration of mice induced with allergic diseases

[0123] The animals used in the experiment were purchased 8-week-old female BALB / c mice (Orient, Korea), which were acclimatized for one week and then used in the experiment at 9-week-old. To induce atopic dermatitis, depilation was performed on the back of BALB / c mice up to the upper part of the back with the largest epilator. 1×1cm on the depilated back at intervals of 24 hours 2 40 μg of Aspergillus fumigatus (Af, Aspergillus fumigatus ) extract was applied to the area of ​​??for 5 days. After a rest period of 2 weeks, the application was repeated 5 times at intervals of 24 hours from the 19th day, thereby completing the preparation of the animal model of allergic dermatitis.

[0124] After the animal model of allergic dermatitis was prepared, MSC, iMSC, MSC-Exo (exosomes derived from mesenchymal stem cells) or iMSC-Exo (mesenchymal stem cells derived from induced pluripotent stem cel...

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Abstract

The present invention may provide a pharmaceutical composition for preventing or treating skin diseases, a cosmetic composition for preventing or improving skin diseases, and a stem cell therapeutic agent, which comprise an induced pluripotent stem cell-derived mesenchymal stem cell (iMSC), a culture thereof, or an exosome isolated from an iMSC or iMSC culture as active ingredients. When the composition of the present invention, etc. is used, it is possible to provide a composition for improving, preventing or treating skin diseases and a stem cell therapeutic agent which have an improved immunomodulating function over conventional mesenchymal stem cells.

Description

technical field [0001] This invention was completed under the support of the Korean Ministry of Science and Technology Information and Communication. The project number is NRF-2015K1A4A3046807. The research management organization of the above-mentioned project is the Korean Research Foundation. "Asan-Minnesota Transplant Innovation Center", the competent authority is "Seoul Asan Hospital", and the research period is from September 01, 2015 to August 31, 2021. [0002] This application claims priority from Korean Patent Application No. 10-2018-0001978 filed with the Korean Patent Office on January 5, 2018, the disclosure of which is hereby incorporated by reference into this specification. [0003] The present invention relates to a medicinal product comprising induced pluripotent stem cell-derived mesenchymal stem cells (iMSCs), cultures thereof, or exosomes (iMSC-exo) isolated from iMSCs or cultures of iMSCs as active ingredients. Composition for improving, preventing or tr...

Claims

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Application Information

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IPC IPC(8): A61K35/28A61K35/545A61K9/127A61K8/98A61K8/14A61P17/00A61Q19/00
CPCA61K8/14A61K9/127A61K35/28A61K35/545A61P17/00A61Q19/00A61K8/99A61K8/981A61P17/10A61K8/982A61K8/983
Inventor 金秀刘珍虎吴然睦
Owner THE ASAN FOUND
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