WO2021225260A1 - Composition de cicatrisation des plaies comprenant un milieu conditionné par des cellules régulatrices t - Google Patents

Composition de cicatrisation des plaies comprenant un milieu conditionné par des cellules régulatrices t Download PDF

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WO2021225260A1
WO2021225260A1 PCT/KR2021/002192 KR2021002192W WO2021225260A1 WO 2021225260 A1 WO2021225260 A1 WO 2021225260A1 KR 2021002192 W KR2021002192 W KR 2021002192W WO 2021225260 A1 WO2021225260 A1 WO 2021225260A1
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cells
regulatory
cell
composition
cell culture
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황성환
강정화
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주식회사 이뮤니스바이오
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/14—Blood; Artificial blood
    • A61K35/17—Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0634—Cells from the blood or the immune system
    • C12N5/0636—T lymphocytes
    • C12N5/0637—Immunosuppressive T lymphocytes, e.g. regulatory T cells or Treg
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/10—Growth factors
    • C12N2501/15—Transforming growth factor beta (TGF-β)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/20—Cytokines; Chemokines
    • C12N2501/23—Interleukins [IL]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/50—Cell markers; Cell surface determinants
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/70—Enzymes
    • C12N2501/71—Oxidoreductases (EC 1.)

Definitions

  • the present invention relates to a regulatory T cell culture medium having a wound healing effect, and more particularly, to a wound healing composition comprising a regulatory T cell culture medium having a wound healing effect obtained by culturing regulatory T cells as an active ingredient.
  • the skin is one of the largest organs in the human body, and its main function is to protect the human body from external damage, temperature changes, and environmental changes.
  • the skin consists of two layers: the epidermis and the dermis.
  • the epidermis is the outermost layer of the skin and is mostly composed of keratinocytes, and cells proliferate in the basal layer and naturally fall off as they move to the outer stratum corneum.
  • the dermis is the inner layer of the skin and is supported by an extracellular matrix (ECM) to provide cushioning and tensile strength of the skin.
  • ECM extracellular matrix
  • a wound is defined as an abnormal condition of the skin caused by causes such as trauma or disease. If the wound persists, the skin cannot perform its full function, so the wound healing process is very important.
  • the wound healing process can be simply divided into hemostasis, inflammation, re-epithelialization, and tissue remodeling.
  • Re-epithelialization is a process in which epithelial cells move to the surface of the wound and cover the wound.
  • keratinocytes of the epidermis migrate to the wound site and are mainly involved.
  • epithelium epithelium
  • keratinocytes go through the processes of migration, differentiation, and proliferation.
  • EMT Epithelial-Mesenchymal Transition, epithelial-mesenchymal transition
  • the EMT process is essential in the wound healing process, and EMT allows mobility, decreases the expression of epithelial cell marker proteins, and increases the expression of mesenchymal cell marker proteins.
  • the EMT process is regulated by matrix metalloproteinases (MMPs), adhesion junction proteins such as E-cadherin, and transcription factors such as Twist and Snail.
  • MMPs matrix metalloproteinases
  • adhesion junction proteins such as E-cadherin
  • transcription factors such as Twist and Snail.
  • MMP Metalloproteinase
  • MMP plays a role in decomposing the extracellular matrix and reduces the adhesion of keratinocytes to the extracellular matrix in the tissue restructuring step, thereby allowing the movement of keratinocytes and restructuring the tissue. let it be In addition, MMP plays an important role in all stages of wound healing by enabling cell migration and tissue restructuring by removing or restructuring epithelial and interstitial extracellular matrix components.
  • the extracellular matrix is degraded by various proteinases, and Type I collagen, which is the most abundant in the skin, is resistant to most enzymes.
  • Matrix metalloproteinase-1 (MMP-1) degrades collagen, particularly Type I collagen, which is most abundantly present in the skin and accounts for the majority of the extracellular matrix.
  • MMP-1 plays an important role in cell signaling by being involved in the secretion of various growth factors and cytokines.
  • the expression of MMP is very complexly regulated, and under normal conditions, it maintains a basal level and is selectively expressed and activated only when tissue restructuring is required.
  • E-cadherin is one of the adhesive junction protein (Adherant junction protein) has the function of maintaining the intercellular junction and epithelial phenotype, and is used as an EMT marker protein to identify EMT.
  • cytoskeleton rearrangement occurs in epithelial cells, cell junction and apical-basal polarity are weakened, and binding to the extracellular matrix is impaired. will change And it acquires mobility, which is one of the mesenchymal features (Thiery et al., 2009; Lim and thiery, 2012; Lamouille et al, 2014; Niero et al., 2016).
  • the adherent junction becomes unstable, resulting in down-regulation of E-cadherin.
  • Immune cells play a very important role in the wound healing process. Immune cells contribute to the removal of foreign antigens when an inflammatory response occurs after a wound, and secrete various growth factors and cytokines in the wound healing process to influence tissue restructuring and cell migration.
  • the immune system is regulated by very complex interactions (cross-talk), and it is very important that the immune system is well regulated in order to maintain body homeostasis.
  • Regulatory T cells are a subpopulation of T cells and play a very important role in regulating the immune response. Regulatory T cells show a self-tolerance effect that regulates the activation of the immune system and prevents autoimmune disease, which is a pathological self-reactivity.
  • regulatory T cells can also affect surrounding cells by secreting various substances such as cytokines and growth factors. Cytokines are a category of signaling molecules that regulate immunity, inflammation, and hematopoiesis. They are essential substances for interaction and information transfer between immune cells and are involved in cell migration, proliferation, and inflammatory responses. . For example, it has been reported that IL-8 significantly increases the migration of keratinocytes.
  • the wound healing process is very complex and finely regulated due to the involvement of various factors. have.
  • Patent Document 1 Korean Patent Publication No. 10-2018-0057359
  • Patent Document 2 Korean Patent Publication No. 10-2019-0076692
  • Patent Document 3 US Patent Publication US2014/0112898
  • Non-Patent Document 1 Immune Cell-Conditioned Media Suppress Prostate Cancer PC-3 Cell Growth Correlating With Decreased Proinf lammatory/Anti-inflammatory Cytokine Ratios in the Media Using 5 Selected Crude Polysaccharides, Hsiao-Chien Lin et al., Integrative Cancer Therapies 2016, Vol. 15(4) NP13-NP25.
  • Non-Patent Document 2 (Non-Patent Document 2) 'Repair' Treg Cells in Tissue Injury, Chaoqi Zhanga et al., Cell Physiol Biochem 2017;43:2155-2169
  • the present invention aims to provide a wound healing composition comprising the same as an active ingredient by confirming that a culture medium (conditioned media) cultured with regulatory T cells, a type of immune cells, has a wound healing effect. do it with
  • the present invention provides a composition for wound healing containing, as an active ingredient, a regulatory T cell culture solution obtained by centrifugation after culturing regulatory T cells.
  • the composition preferably contains 1 to 10% by weight of the regulatory T cell culture solution in the composition.
  • the regulatory T cell culture medium In the composition, the regulatory T cell culture medium,
  • PBMCs from the blood
  • CD8+ cells from the isolated PBMC
  • obtaining a culture solution by culturing for 12 to 16 days while subculturing at intervals of 2-3 days after the incubation and centrifuging the culture solution to remove regulatory T cells and obtain a regulatory T cell culture solution.
  • composition it is preferable to use a phenol red-free and antibiotic-free medium as the cell culture medium.
  • said medium is anti-CD2, anti-CD3, anti-CD7, anti-CD8, anti-CD28, anti-CD30L, anti-CD40, anti-CD70, anti-CD80, anti-CD83 and anti-CD86 at least one antibody selected from the group consisting of; one or more cytokines selected from the group consisting of interleukin-2, interleukin-4, interleukin-7, interleukin-12, interleukin-15, interleukin-34 and TGF- ⁇ ; and SOD (superoxide dismutase) may be further included.
  • cytokines selected from the group consisting of interleukin-2, interleukin-4, interleukin-7, interleukin-12, interleukin-15, interleukin-34 and TGF- ⁇
  • SOD superoxide dismutase
  • the CD8+ cells are preferably removed using a MS column by mixing the cell suspension in MACS buffer and CD8 microbeads.
  • the PBMCs are preferably cultured at a concentration of 1.0 ⁇ 10 7 to 3.0 ⁇ 10 7 cells/flask.
  • the subculture is preferably performed while adding a medium or expanding the flask area so that the cell concentration is 0.5 ⁇ 10 6 to 2.0 ⁇ 10 6 cells/mL.
  • the centrifugation is preferably carried out at 100 ⁇ 1,000 ⁇ g.
  • the present invention also provides a composition for soothing sensitive skin and alleviating atopic dermatitis through wound healing, comprising as an active ingredient a regulatory T cell culture medium obtained by culturing regulatory T cells and then centrifuging.
  • the regulatory T cell culture of the present invention is IL-6. It contains various cytokines and growth factors such as IL-8 and IL-13, and dose-dependently reduces the expression of E-cadherin, a marker protein that identifies EMT, to promote EMT, which is essential in cell movement. It increases the cell migration of keratinocytes without affecting the cell proliferation because it makes the cells have mobility, and increases the expression and release of MMP-1, which affects cell migration and the decomposition of the extracellular matrix.
  • E-cadherin a marker protein that identifies EMT
  • composition comprising the regulatory T cell culture medium of the present invention as an active ingredient can be usefully used for wound healing, and can also be usefully used for soothing sensitive skin and alleviating atopy through wound healing.
  • Figure 2 is the result of analyzing the cytokines in the regulatory T cell culture.
  • the present invention relates to a composition for wound healing containing a regulatory T cell culture medium obtained by centrifugation after culturing regulatory T cells as an active ingredient.
  • PBMCs from the blood
  • CD8+ cells from the isolated PBMC
  • the present invention also relates to a composition for soothing sensitive skin and alleviating atopic dermatitis through wound healing, comprising, as an active ingredient, a regulatory T cell culture medium obtained by culturing regulatory T cells and then centrifuging.
  • PBMC peripheral blood mononuclear cells
  • CD8+ cells are preferably removed by mixing the cell suspension in MACS buffer with CD8 micro beads and using an MS column (Miltenyi Biotec).
  • the cell suspension in MACS buffer and CD8 microbeads are preferably mixed in a ratio (v/v) of 4:1.
  • PBMCs from which CD8+ cells have been removed are cultured in a medium at 37° C., 5% CO 2 conditions for 1 to 3 days, more preferably for 2 days.
  • a phenol red free and antibiotics free medium as the cell culture medium.
  • MEM Minimum Essential Medium
  • DMEM Dulbecco's Modified Eagle Medium
  • BME Basic Medium Eagle
  • RPMI MEM- ⁇ (Minimal Essential Medium- ⁇ )
  • IMDM Iscove's Modified Dulbecco's
  • MacCoy's 5A medium Swim's S-77 medium
  • CTS OpTmizer Endothelial Basal Medium (EBM) medium
  • X-Vivo medium may include one or more medium selected from the group consisting of, but is not limited thereto.
  • interleukins promote the development, maintenance, survival and expansion of regulatory T cells
  • anti-CD antibodies are important costimulatory molecules and modulators of regulatory T cells
  • SOD plays a role in cell survival and survival when present in excess. This is because it improves the reduced cell survival and cell differentiation ability by removing ROS that affects differentiation.
  • PBMCs are preferably cultured at a concentration of 1.0 ⁇ 10 7 to 3.0 ⁇ 10 7 cells/flask, more preferably at a concentration of 2.0 ⁇ 10 7 cells/flask.
  • the culture solution is obtained by culturing for 12 to 16 days while subcultured at intervals of 2 to 3 days. Incubation for 14 days is particularly preferred.
  • the subculture is performed while adding a medium or increasing the flask area so that the cell concentration is 0.5 ⁇ 10 6 to 2.0 ⁇ 10 6 cells/mL.
  • the culture medium in which the regulatory T cells are cultured is centrifuged to remove the regulatory T cells, and a Treg cell-conditioned media is obtained.
  • the degree of CD4+CD25+ regulatory T differentiation is checked, and when 70% or more of the regulatory T differentiation degree (Position) is confirmed, centrifugation is preferable.
  • Centrifugation is preferably centrifuged at 100 to 1,000 ⁇ g, more preferably centrifuged to 300 to 700 ⁇ g.
  • the composition of the present invention preferably contains 1 to 10% by weight of the regulatory T cell culture solution, more preferably 1 to 5% by weight, and particularly preferably 5%.
  • the wound healing effect is excellent when the regulatory T cell culture medium of the above range is included. In particular, when 5% by weight of the regulatory T cell culture medium was included, the cell viability decreased slightly without significant difference, but the wound closure rate was confirmed to be the maximum.
  • Regulatory T cell culture medium of the present invention is IL-6. It contains various cytokines and growth factors such as IL-8 and IL-13, and dose-dependently reduces the expression of E-cadherin, a marker protein that identifies EMT, to promote EMT, which is essential in cell movement. This increases the cell migration of keratinocytes without affecting the cell proliferation by making the cells have mobility, and increases the expression and release of MMP-1, thereby affecting cell migration and decomposition of the extracellular matrix.
  • IL-6 contains various cytokines and growth factors such as IL-8 and IL-13, and dose-dependently reduces the expression of E-cadherin, a marker protein that identifies EMT, to promote EMT, which is essential in cell movement. This increases the cell migration of keratinocytes without affecting the cell proliferation by making the cells have mobility, and increases the expression and release of MMP-1, thereby affecting cell migration and decomposition of the extracellular matrix.
  • the regulatory T cell culture medium of the present invention has excellent wound healing effect, and the composition containing the regulatory T cell culture medium of the present invention as an active ingredient can be usefully used for wound healing, soothing sensitive skin through wound healing effect and It can also be usefully used for alleviating atopy.
  • DMEM Dulbecco's modified Eagle's medium
  • penicillin solution penicillin solution
  • streptomycin solution fetal bovine serum
  • BCA bisinchoninic acid
  • FITC-conjugated secondary antibody fluorescein isothiocyanate-conjugated secondary antibody
  • SDS sodium dodecyl sulfate
  • DMSO dimethyl sulfoxide
  • GPDH glyceraldehyde-3-phosphate dehydrogenase
  • E-cadherin MMP-1 antibody
  • ECL Enhanced chemiluminescence
  • HaCaT cells (ATCC 12192) (Spontaneously immortalized human keratinocyte line) were cultured in DMEM (full medium) supplemented with 10% (v/v) FBS, 100 U/L penicillin, and 100 ⁇ g/mL streptomycin at 37° C., 5% CO 2 cultured under conditions. Serum starvation was performed before HaCaT cells were treated with regulatory T cell culture medium.
  • PBMCs were isolated from blood, and the separated PBMCs were washed with MACS buffer. After mixing the cell suspension in MACS buffer and CD8 microbeads at a ratio (v/v) of 4:1, PBMCs from which CD8+ cells were removed were obtained using an MS column.
  • PBMCs from which CD8+ cells were removed were cultured in a phenol red, antibiotic-free medium at 37° C. and 5% CO 2 conditions. As the cells proliferated, it was subcultured while adding the medium to an appropriate cell concentration. After culturing for 14 days, the culture medium in which the regulatory T cells were cultured was centrifuged to obtain a regulatory T cell culture in which the regulatory T cells were removed.
  • the regulatory T cells isolated by centrifugation in Example 1 were collected, washed, and centrifuged. After centrifugation, the cells were resuspended in ice cold FACS buffer, and cultured in the presence of saturated CD4 and CD25 antibodies to stain regulatory T cells. The surface expression of CD4 and CD25 molecules was confirmed by flow cytometry of the stained regulatory T cells.
  • Foxp3 X chromosome-encoded forkhead transcription factor
  • FIG. 1 The results of confirming the surface expression of CD4 and CD25 molecules and the expression of Foxp3 by flow cytometry analysis of regulatory T cells isolated from PBMCs from which CD8+ cells were removed are shown in FIG. 1 .
  • regulatory T cells expressed CD4 and CD25 but not CD8, and CD4+CD25+ regulatory T cells from which CD8+ cells were removed were isolated from PBMC by 90.6%.
  • Cytokine analysis was performed to confirm the cytokines contained in the culture medium in which the regulatory T cells were cultured.
  • each cytokine array membrane After blocking each cytokine array membrane, it was reacted with a regulatory T cell culture medium. Controls were reacted with medium alone. It was washed with a wash buffer and reacted with a biotinylated antibody cocktail. After washing, the reaction was performed using HRP-streptavidin solution. After the reaction was completed, the membrane was washed and reacted with a detection mixture buffer, followed by detection using chemidoc.
  • FIG. 2 (A) The results of detecting the regulatory T cell culture by Chemidoc are shown in FIG. 2 (A), and the results of confirming the relative strength of each cytokine by densitometric analysis are shown in FIG. 2 (B).
  • FIG. 2 (A) the left image shows the results when the medium alone was used as a control, the right image shows the results of the regulatory T cell culture, and the table shows the cytokine position of each dot.
  • IL-6, IL-8, IL-10, IP-10, MCP-1, and RANTES were significantly increased. It has been reported that IL-6, IL-8, IL-10, IP-10, MCP-1, and RANTES all increased cell migration, especially of keratinocytes. Through cytokine analysis, it can be confirmed that IL-6 increased about 70 times than the basal level, and IL-8, IL-10, IP-10, MCP-1, and RANTES increased about 40 times.
  • MTT analysis was performed to determine the effect of regulatory T cell culture on the proliferation of HaCaT keratinocytes.
  • HaCaT cells were cultured in a 96-well culture plate, the cells were treated with various concentrations of control T cell culture medium, and then cell viability was confirmed using the MTT assay.
  • MTT reagent was treated and cultured in each well, and then the generated formazan was dissolved in DMSO and absorbance was measured at 560 nm. The results were calculated as the mean ⁇ standard deviation of the 6th experiment and shown in FIG. 3 .
  • cells treated with the regulatory T cell culture medium for 24 hours showed a slight decrease in cell proliferation compared to control cells that were not treated. That is, when the regulatory T cell culture was treated at a concentration of 5%, it was reduced by about 5%, and when treated with 20%, it was confirmed to be reduced by about 20%.
  • the regulatory T cell culture does not promote the proliferation of HaCaT cells.
  • FIG. 4 The results are shown in FIG. 4 .
  • Figure 4 (A) shows the microscopic image obtained immediately after 24 hours for the scratched cell monolayer
  • (B) shows the result of confirming cell migration by measuring the decrease in the scratched area. The results are expressed as the mean ⁇ standard deviation of the 6th experiment.
  • the cells treated with the regulatory T cell culture medium had a reduced scratch area compared to the untreated control, and cell migration was increased when the regulatory T cell culture medium was pretreated. From these results, it can be seen that the regulatory T cell culture of the present invention can affect the re-epithelialization and regulation of the wound site.
  • E-cadherin a marker protein of EMT
  • FITC fluorescence images were obtained using a LEICA DMi8 fluorescence microscope (Leica microsystems, Wetzlar, Germany).
  • Figure 5 (A) is the result of detecting E-cadherin by immunofluorescence staining, (B) is the result of confirming the relative intensity by densitometric analysis.
  • E-cadherin in HaCaT cells pretreated with regulatory T cell culture was significantly reduced in a dose-dependent manner. From these results, it can be confirmed that the regulatory T cell culture medium promotes EMT at the wound site, thereby promoting the movement of keratinocytes and down-regulating E-cadherin.
  • HaCaT keratinocytes were treated with RIPA (radioimmunoprecipitation assay) lysis buffer containing a protease/phosphatase inhibitor mixture to obtain a whole cell lysate, and then the protein concentration was measured with a BCA reagent.
  • RIPA radioimmunoprecipitation assay
  • the medium protein or lysate protein was separated by SDS-PAGE (SDS-polyacrylamide gel electrophoresis) gel and transferred to a polyvinylidene fluoride membrane.
  • the protein-transferred membrane was treated with skim milk for non-specific blocking.
  • Blot (Blot), after reacting the primary antibody diluted 1/4000, Horseradish peroxidase-conjugated secondary antibody (Horseradish peroxidase-conjugated secondary antibody) was detected using ECL reagent. Bands were quantified with a densitometer.
  • FIG. 6 The results of confirming the effect of regulatory T cell culture on MMP-1 expression and secretion in HaCaT keratinocytes are shown in FIG. 6 .
  • GAPDH was used as a loading control for MMP-1.
  • (A) is the result of analyzing the total cell lysate (L) and the medium (M) MMP-1 by Western blotting
  • (B) is the result of confirming the relative intensity of MMP-1 by densitometric analysis.
  • the results of FIG. 6 show that treatment with regulatory T cell culture for 24 hours significantly increased both the expression and secretion of MMP-1 in a dose-dependent manner. Therefore, it is confirmed that ECM degradation and restructuring at the wound site are activated and the expression of MMP-1 is reduced when the regulatory T cell culture medium is treated.

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Abstract

La présente invention se rapporte à une composition de cicatrisation des plaies, la composition contenant, comme principe actif, un milieu conditionné par des cellules Treg obtenu par culture des cellules T régulatrices (cellules Treg) et ensuite leur centrifugation. Le milieu conditionné par des cellules Treg selon la présente invention contient diverses cytokines, telles que l'IL-6, l'IL-8, ou l'IL-13, et des facteurs de croissance, elle réduit en fonction de la dose l'expression de la E-cadhérine, qui est une protéine marqueur identifiant la transition épithéliale-mésenchymateuse (EMT) qui se produit nécessairement durant la migration des cellules pour favoriser l'EMT, donnant ainsi aux cellules la capacité de migrer et augmentant la migration des kératinocytes sans affecter la prolifération des cellules, et qui accroît l'expression et la libération de la métalloprotéinase de la matrice 1 (MMP-1), affectant ainsi la migration des cellules et la décomposition de la matrice extracellulaire. Ainsi, la composition contenant le milieu conditionné par des cellules Treg comme principe actif selon la présente invention peut être utile pour permettre la cicatrisation des plaies, ainsi que pour calmer la peau sensible et soulager la dermatite atopique à travers la cicatrisation des plaies.
PCT/KR2021/002192 2020-05-04 2021-02-22 Composition de cicatrisation des plaies comprenant un milieu conditionné par des cellules régulatrices t WO2021225260A1 (fr)

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Cited By (1)

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Publication number Priority date Publication date Assignee Title
CN115737543A (zh) * 2022-12-12 2023-03-07 中南大学 调节性t细胞囊泡的制备方法、复合水凝胶及其应用

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