Application of adipose-derived stem cell exosome in hair growth

An adipose stem cell and exosome technology, which is applied in the field of medical use of stem cell exosomes, can solve the problems of difficulty in completely separating non-vesicular entities and extracellular vesicle components, affecting the composition and functional differences of extracellular vesicles, etc. , to achieve increased number and dermal thickness, low immunogenicity, easy storage

Pending Publication Date: 2022-04-29
SHANGHAI TONGJI HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, we know that the extracellular environmental components are complex, and it is difficult for common exosome isolation methods to completely separate non-vesicular entities and extracellular vesicle components. Therefore, the exosome isolation method affects the composition of the isolated extracellular vesicles. and the resulting functional difference
There is no report on mouse adipose stem cell exosomes and their use for hair growth as described in this application

Method used

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  • Application of adipose-derived stem cell exosome in hair growth
  • Application of adipose-derived stem cell exosome in hair growth
  • Application of adipose-derived stem cell exosome in hair growth

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] 1. Materials and methods

[0039] 1 material

[0040] 1.1 Animal source

[0041] SPF-grade C57BL / 6 male mice were used in the experiment, which were purchased from Shanghai Slack Experimental Animal Co., Ltd. and bred in the Experimental Animal Center of Tongji University. The experimental research ethics were approved by the Experimental Animal Center of Tongji University.

[0042] 1.2 Main experimental reagents / consumables

[0043] DMEM medium (Gibco, U.S., article number: C11995500BT), phosphate buffer solution (Phosphate BufferSolution, PBS, Gibco, U.S.), 0.05% trypsin solution (Trypsin-EDTA, Gibco, U.S.), FetalBlood Serum (FBS, Gibco, U.S.), USA, Cat. No.: 10099141C), type I collagenase (Worthington, USA). Ultra-15 Centrifugal Filter Unit (Millipore, USA, Cat. No. UFC910008), ExoQuick-TC (Cat. No. EXOTC10A-1, SBI Biosciences, SBI, America), Anti-CD9 Antibody (abcam, UK), Anti-CD81 Antibody (abcam, UK ), Anti-TSG101 antibody (abcam, UK).

[0044] 2 methods

[...

Embodiment 2

[0068] 1. Materials and methods

[0069] 1 material

[0070] 1.1 Animal source

[0071] SPF-grade C57BL / 6 male mice and SPF-grade SD male rats were used in the experiment, both purchased from Shanghai Slack Experimental Animal Co., Ltd. and bred in the Experimental Animal Center of Tongji University. Experimental research ethics were approved by the Experimental Animal Center of Tongji University.

[0072] 1.2 Main experimental reagents / consumables

[0073] DMEM medium (Gibco, U.S., article number: C11995500BT), phosphate buffer solution (Phosphate BufferSolution, PBS, Gibco, U.S.), 0.05% trypsin solution (Trypsin-EDTA, Gibco, U.S.), FetalBlood Serum (FBS, Gibco, U.S.), USA, product number: 10099141C), type I collagenase (Worthington, USA), CCK8 reagent (Cell Counting Kit-8, Dojindo, Japan).

[0074] 2 methods

[0075] 2.1 Exosomes from adipose-derived stem cells promote the proliferation of hair follicles and dermal papilla

[0076] The 3rd-5th generation rat hair follic...

Embodiment 3

[0085] 1. Materials and methods

[0086] 1 material

[0087] 1.1 Animal source

[0088] SPF-grade C57BL / 6 male mice and SPF-grade SD male rats were used in the experiment, both purchased from Shanghai Slack Experimental Animal Co., Ltd. and bred in the Experimental Animal Center of Tongji University. Experimental research ethics were approved by the Experimental Animal Center of Tongji University.

[0089] 1.2 Main experimental reagents / consumables

[0090] DMEM medium (Gibco, U.S., article number: C11995500BT), phosphate buffer solution (Phosphate BufferSolution, PBS, Gibco, U.S.), 0.05% trypsin solution (Trypsin-EDTA, Gibco, U.S.), FetalBlood Serum (FBS, Gibco, U.S.), USA, Cat. No.: 10099141C), type I collagenase (Worthington, USA). Ultra-15 Centrifugal Filter Unit (Millipore, USA, article number UFC910008), ExoQuick-TC (article number EXOTC10A-1, SBI Biosciences, SBI, the United States). CCK8 reagent (Cell Counting Kit-8, Dojindo, Japan).

[0091] 2 methods

[0092] ...

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Abstract

A preparation method of the adipose-derived stem cell exosome comprises the following steps: culturing 3rd to 6th generations of mouse adipose-derived stem cells in logarithmic growth by using a complete culture solution containing FBS until the cell density reaches 70 to 80 percent, and continuously culturing for 45 to 50 hours by using a DMEM culture solution containing exosome-free FBS, so as to obtain the adipose-derived stem cell exosome. And collecting a supernatant culture solution, centrifuging to remove cells and cell debris, centrifuging at 1500-2500g by using an ultrafiltration tube with the molecular weight of 100kDa for 30 minutes, concentrating, adding an exosome rapid extraction reagent into a concentrated solution, extracting, centrifuging, and collecting the mouse adipose-derived stem cell exosome. In-vitro and in-vivo experiments show that the mouse adipose-derived stem cell exosome can significantly promote hair growth and has an outstanding effect.

Description

technical field [0001] The present invention relates to the medical use of stem cell exosomes, in particular to the use of fat stem cell exosomes in hair growth. Background technique [0002] Adipose-derived stem cells (ADSCs) are a type of mesenchymal stem cells that exist in adipose tissue and have multilineage differentiation potential. In vitro and in vivo, ADSCs have the potential to differentiate into adipocytes, chondrocytes, osteoblasts, neurons, vascular endothelial cells, tenocytes and other cell lines. [0003] Exosomes are a type of extracellular vesicles with a lipid bilayer structure and a diameter of 30-150 nm that can be secreted by cells into the cell culture supernatant. Exosomes contain a variety of proteins, nucleic acids (miRNA, mRNA, lncRNA, circRNA) and lipids. They are important carriers of intercellular communication, and almost all cells can release exosomes. [0004] Exosomes are widely used and can be used for the treatment of various diseases, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775A61K35/28A61P17/14
CPCC12N5/0667A61K35/28A61P17/14C12N2509/00
Inventor 黄欣李燕巧
Owner SHANGHAI TONGJI HOSPITAL
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