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Composition for improving, preventing or treating skin disease comprising induced pluripotency stem cell-derived mesenchymal stem cells pretreated with interferon gamma and exosomes derived therefrom

A technology of pluripotent stem cells and mesenchymal stem cells, applied in artificially induced pluripotent cells, skin diseases, skin care preparations, etc., can solve problems such as no known treatment methods

Pending Publication Date: 2020-08-28
THE ASAN FOUND +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For atopic dermatitis, there is no known direct specific treatment so far, and 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 disease comprising induced pluripotency stem cell-derived mesenchymal stem cells pretreated with interferon gamma and exosomes derived therefrom
  • Composition for improving, preventing or treating skin disease comprising induced pluripotency stem cell-derived mesenchymal stem cells pretreated with interferon gamma and exosomes derived therefrom
  • Composition for improving, preventing or treating skin disease comprising induced pluripotency stem cell-derived mesenchymal stem cells pretreated with interferon gamma and exosomes 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 the absence of adipocytes (cultivated helper cells), a serum supplement containing KnockOut gene-free serum replacement, glutamine, non-essential amino acids, β-mercaptoethanol, antibiotics, and basic fibroblast growth factor (bFGF, Induced pluripotent stem cells cultured in Dulbecco's modified Eagle's medium / F-12 medium (basicfibroblast growth factor) (derived from fibroblasts, peripheral blood established in Mononuclear cells (Peripheral Blood Mononuclear Cell, PBMC), induced pluripotent stem cell (iPSC) cell line of mesenchymal stem cells) were attached to the culture dish (dish) coated with vitronectin in advance, and then added with 10% Fetal bovine serum (FBS) (v / v), 5ng / ml basic fibroblast growth factor (basic FGF), 0.1mM minimum essential medium non-essential amino acids (MEM NEAA, Minimum Essential Media Non-Essential AminoAcids) , β-mercaptoet...

Embodiment 2

[0118]Example 2: Isolation and confirmation of exosomes derived from mesenchymal stem cells differentiated from induced pluripotent stem cells according to whether interferon-γ pretreatment was used

[0119] Induced pluripotent stem cell-derived mesenchymal stem cells and induced pluripotent stem cell-derived mesenchymal stem cells pretreated with interferon-γ were further cultured in a medium supplemented with 10% exosome-depleted fetal bovine serum (Exosome-depleted FBS). stem cells. After culturing for 72 hours, the induced pluripotent stem cell-derived mesenchymal stem cell culture medium was collected and centrifuged at 300×g for 10 minutes to remove excess cells and cell residues. The supernatant was filtered through a 0.22 μm filter, and then centrifuged at 10,000×g and 4° C. for 70 minutes using a high-speed centrifuge. Centrifuge the centrifuged supernatant at 100,000xg and 4°C for 90 minutes in an ultracentrifuge to remove the supernatant, and dilute the exosomes re...

Embodiment 3

[0122] Example 3: Preparation and Administration of Mice Inducing Specific Diseases

[0123] Animals used in the experiment were purchased 8-week-old female BALB / c mice (Orientbio Inc., Korea), and used for the experiment at 9-week-old after a one-week acclimation period. In order to induce atopic dermatitis, BALB / c mice were shaved up to the upper dorsal side with a shaving machine. in 1×1cm 2 40 μg of Aspergillus fumigatus (Af, Aspergillus fumigatus) extract was applied for a total of 5 days at intervals of 24 hours. After a 2-week rest period, the application was repeated 5 times at 24-hour intervals on the 19th day to prepare an atopic dermatitis animal model.

[0124] After the animal model of atopic dermatitis was prepared, mesenchymal stem cells derived from induced pluripotent stem cells, mesenchymal stem cells derived from induced pluripotent stem cells pretreated with interferon γ, and Exosomes from mesenchymal stem cells and exosomes from induced pluripotent stem...

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Abstract

The present invention may provide a method for producing induced pluripotency stem cell-derived mesenchymal stem cells (IFN gamma-iMSC) pretreated with interferon-gamma; induced pluripotency stem cell-derived mesenchymal stem cells pretreated with interferon-gamma prepared by the method; a pharmaceutical composition for preventing or treating skin diseases, and a cosmetic composition for preventing or improving skin diseases, comprising culture thereof or exosome (IFN gamma-iMSC-exo) isolated therefrom as an active ingredient. When the composition of the present invention is used, it is possible to provide a composition for 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 also completed under the support of the Korean Ministry of Science and Technology Information and Communication through the subject number NRF-2015K1A4A3046807. The research management agency for the above-mentioned subject is the Korean Research Foundation. "Asan-Minnesota Transplantation Innovation Center", the competent institution is "Seoul Asan Hospital", and the research time is from September 01, 2015 to August 31, 2021. [0002] This application claims priority from Korean Patent Application No. 10-2018-0001979 filed with the Korean Patent Office on January 5, 2018, the disclosure of which is incorporated herein by reference. [0003] The present invention relates to induced pluripotent stem cell-derived mesenchymal stem cells (IFNγ-iMSC) comprising interferon gamma pretreatment, culture thereof and / or induced pluripotent stem cell-derived mesenchymal stem cells derived from interferon gamma pretreatment Compositions for impro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775A61K8/98A61K35/28A61P17/00A61P17/06A61Q19/00
CPCA61K8/98A61K35/28A61P17/00A61P17/06A61Q19/00C12N5/0662C12N2501/24C12N2506/45A61K35/12A61K8/981C12N2533/90C12N5/0652
Inventor 金秀刘珍虎
Owner THE ASAN FOUND
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