Use of composition comprising stem cell-derived exosome as effective ingredient in reinforcement and functional improvement of skin barrier

A skin barrier and composition technology, applied in the field of skin external preparations and cosmetic compositions, can solve complex and expensive, anti-host disease and other problems, to overcome safety problems, or improve skin barrier function, enhance skin barrier or improve skin The effect of barrier function

Pending Publication Date: 2020-07-17
韩国外泌体生技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the use of allogeneic adult stem cells (not the patient's own) poses a risk of developing graft-versus-host disease
When using autologous adult stem cells for therapy, a problem arises: the process of culturing adult stem cells isolated from the patient is necessary, which is complex and expensive
Therefore, when conditioned medium is applied to the skin, the skin is highly likely to be exposed to several risks

Method used

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  • Use of composition comprising stem cell-derived exosome as effective ingredient in reinforcement and functional improvement of skin barrier
  • Use of composition comprising stem cell-derived exosome as effective ingredient in reinforcement and functional improvement of skin barrier
  • Use of composition comprising stem cell-derived exosome as effective ingredient in reinforcement and functional improvement of skin barrier

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] Example 1: Cell Culture

[0085] RAW 264.7 cells (mouse macrophage cell line) were purchased from Korean Cell Line Bank and cultured. For cell culture, cells were grown in DMEM (available from ThermoFisher Scientific) medium containing 10% fetal bovine serum (FBS; available from ThermoFisher Scientific) and 1% antibiotic-antimycotic (available from ThermoFisher Scientific) At 37°C, 5% CO 2 Under subculture.

[0086] Human dermal fibroblast (HDF) HS68 cells purchased from ATCC were prepared in DMEM (purchased from ThermoFisher Scientific) containing 10% fetal bovine serum (FBS; purchased from ThermoFisher Scientific) and 1% antibiotic-antimycotic agent (purchased from ThermoFisher Scientific). Scientific) culture medium at 37°C, 5% CO 2 Under subculture.

[0087] According to the cell culture method known in the technical field of the present invention, at 37 ℃, 5% CO 2 cultured adipose-derived stem cells. Next, the cells were washed with phosphate-buffered saline (p...

Embodiment 2

[0089] Example 2: Exosome Isolation and Purification by the TFF Method

[0090] For the isolation, concentration and diafiltration of exosomes from the conditioned medium in Example 1 filtered through a 0.22 μm filter, the TFF method was used. As a filter for the TFF method, a cartridge filter (also called a hollow fiber filter; available from GE Healthcare) or a cassette filter (available from Pall, Sartorius or Merck Millipore) was used. TFF filters with different molecular weight cut-offs (MWCO) are available. Using a filter with the selected MWCO, exosomes are separated and concentrated, and particles smaller than the MWCO, proteins, lipids, nucleic acids, low molecular weight compounds, etc. are removed.

[0091] For separation and concentration of exosomes, TFF filters with MWCO of 100,000 Da (Daltons), 300,000 Da or 500,000 Da were used. Exosomes are isolated from conditioned media by the TFF method by removing material smaller than the MWCO and concentrating the cond...

Embodiment 3

[0093] Example 3: Characterization of Isolated Exosomes

[0094] Particle size and concentration of isolated exosomes were measured by Nanoparticle Tracking Analysis (NTA) (from Malvern) or Tunable Resistance Pulse Sensing (TRPS) (from Izon Science). The homogeneity and size of isolated exosomes were analyzed using transmission electron microscopy (TEM). The results of TRPS, NTA and TEM analysis of isolated exosomes according to one embodiment of the present invention are shown in Figures 1A to 1C.

[0095] Figure ID shows the results of Western blot analysis of exosomes isolated by the isolation method according to one embodiment of the present invention. As indicated therein, the presence of CD9, CD63, CD81 and TSG101 markers was determined. As antibodies against each marker, anti-CD9 (purchased from Abcam), anti-CD63 (purchased from System Biosciences), anti-CD81 (purchased from System Biosciences) and anti-TSG101 (purchased from Abcam) were used, respectively.

[0096] ...

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Abstract

A composition for reinforcement and functional improvement of the skin barrier according to the present invention enhances objective indices associated with the protection, reinforcement, and functional improvement of the skin barrier. The composition for reinforcement and functional improvement of the skin barrier according to the present invention can improve the water loss index of the stratumcorneum, such as transepidermal water loss (TEWL), and increase skin hydration. In addition, the composition for reinforcement and functional improvement of the skin barrier of the present invention exhibits the effects of increasing contents of ceramide, dihydroceramide, and sphingoid bases, increasing the activity of an enzyme involved in the synthesis thereof, and decreasing the activity of anenzyme involved in the degradation thereof. Furthermore, the composition for reinforcement or functional improvement of the skin barrier of the present invention reduces TSLP, IL-4, and IL-13, which are closely relevant to skin barrier damage, to block the vicious circle of reducing lipids and proteins contributing to the skin barrier and thus can make a contribution to the recovery of skin barrier functions.

Description

technical field [0001] The present invention relates to the use of a composition comprising stem cell-derived exosomes as an active ingredient in strengthening the skin barrier or improving the skin barrier function. [0002] Furthermore, the present invention relates to a pharmaceutical composition, an external preparation for skin, and a cosmetic composition comprising the above composition for enhancing the skin barrier or improving the skin barrier function. Background technique [0003] Skin is usually composed of three layers: epidermis, dermis, and subcutaneous fatty tissue. The epidermis is divided into stratum corneum, translucent layer, granular layer and basal layer from outside to inside. The stratum comeum is the outermost layer of the epidermis, a structure that plays the most important role in the skin barrier function, and is composed of enucleated flat corneocytes and stratum corneum (SC) intercellular lipids. Various types of lipids such as sphingolipids,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K8/98A23L33/10A61Q19/00
CPCA61Q19/00A61K8/14A61K36/82A61K36/9068A61K36/484A61K36/718A61K36/42A61K36/28A61K36/53A61K36/752A61K2300/00A61K8/981A23L33/10A61K2800/80A61K2800/82A23V2002/00A23V2200/318A61K8/0212A61K8/0216A61K8/98A61K9/0019A61K9/06A61K35/12A61K2800/884
Inventor 李龙源赵柄成
Owner 韩国外泌体生技有限公司
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