KR20190044242A - Composition for enhancing the action for hair growth of adipose stem cell comprising Minoxidil as an active ingredient - Google Patents

Abstract

The present invention relates to a composition for enhancing hair growth capacity of adipose stem cells, comprising minoxidil as an effective component. More specifically, the present invention relates to a composition for enhancing hair growth capacity of adipose stem cells, comprising minoxidil as an effective component; a method of enhancing hair growth capacity of adipose stem cells by using the same; and a pharmaceutical composition, a quasi-drug composition, and a cosmetic composition for preventing and treating hair loss or promoting hair growth, comprising adipose stem cells with hair growth capacity enhanced by the method. According to the present invention, the minoxidil-treated adipose stem cells have significantly enhanced reproductive integrity and migratory ability, have increased secretion of growth factors, and when administered in an animal model, serve to significantly induce and increase hair growth in growth period compared to a control group. Accordingly, the minoxidil-treated adipose stem cells according to the present invention may be beneficially used in compositions for preventing hair loss, treating hair loss or atrichia, and improving and promoting hair growth and, more specifically, may be beneficially used as drug compositions, quasi-drug compositions, or cosmetic compositions for preventing hair loss or improving hair growth, comprising the minoxidil-treated adipose stem cells or a culture fluid thereof as an effective component.

Description

TECHNICAL FIELD The present invention relates to a composition for enhancing the excipient activity of adipose stem cells comprising minoxidil as an active ingredient,

The present invention relates to a composition for enhancing the excipient function of adipose stem cells comprising minoxidil as an active ingredient, and more particularly, to a composition for enhancing excipient function of adipose stem cells containing minoxidil as an active ingredient, A method for promoting hair growth, and a method for promoting hair growth, hair growth promoting method, and a cosmetic composition.

As interest in beauty has increased recently, interest in hair loss treatment is also growing. Currently, the most commonly used method for hair loss treatment is hair transplantation using a hair transplant using its own hair, and drug therapy using minoxidil and propecia.

In the case of minoxidil, hair growth is promoted by increasing nutrient supply to the hair cells through vasodilation and potassium channel opening effect. In the case of propecia, hair growth is induced by the effect of inhibiting the production of dihydrotestosterone (DHT) . Both of these drugs show hair growth after treatment, but when treatment is stopped, hair loss resumes, hair loss prevention effect is greater than hair growth effect, and especially side effects such as sexual dysfunction and birth defects in pregnant women There is a problem that it can appear.

In recent years, there has been developed a gene therapy method that is performed by transferring genes related to hair loss to hair follicles or blocking gene expression. However, there is a problem that the efficacy and safety of the treatment are uncertain, and the treatment cost is high and clinical application is not easy. For this reason, besides gene therapy, a method of treating hair loss using stem cells has recently become popular.

Adipose-derived Stem Cells are mesenchymal stem cells, which exhibit skin wound healing and anti-aging effects (Kim et al., J Dermatol Sci 53: 96-102, 2009; Altman et al., Stem Cells 27: 250-258, 2009). Recent studies have shown that mice with a hair cycle that is hypoxic or pre-treated with vitamin C convert telogen to anagen (Won et al., J Dermatol Sci 57: 134-137, 2010; Kim et al., Stem Cells Dev 23: 1364-1376, 2014), which is attributed to the enhancement of hair regeneration ability of adipose stem cells. It is also known that the growth factor PDGF-A, expressed and secreted in adipose stem cells, regulates the activity of follicular stem cells and induces anagen in the hair cycle (Festa et al., Cell 146: 761-771, 2011, Tomita et al., J Dermatol Sci 43: 105-115, 2006). However, the functions and effects of minoxidil in adipose stem cells have not been confirmed yet.

The present inventors have intensively studied to develop a method for improving production yield and stem cell growth of adipose stem cells, and the present inventors have found that the proliferation, migration and growth factor secretion of adipose stem cells are increased by minoxidil treatment, And completed the present invention.

It is an object of the present invention to provide a composition for promoting the production of lipid stem cells containing minoxidil as an active ingredient.

Yet another object of the present invention is to provide a method for promoting the function of lipid stem cells, comprising culturing adipose stem cells in a medium containing minoxidil as an active ingredient.

Yet another object of the present invention is to provide a pharmaceutical composition for preventing, treating or promoting hair loss comprising minoxidil-treated adipose stem cells or a culture thereof as an active ingredient.

It is still another object of the present invention to provide a quasi-drug composition for preventing hair loss or improving hair growth comprising minoxidil-treated adipose stem cells or a culture thereof as an active ingredient.

It is still another object of the present invention to provide a cosmetic composition for preventing hair loss or improving hair growth comprising minoxidil-treated adipose stem cells or a culture thereof as an active ingredient.

Disclosure of Invention Technical Problem [8] Accordingly, the present invention provides a composition for enhancing the excipient function of adipose stem cells comprising Minoxidil as an active ingredient.

In the present invention, adipose-derived stem cells refer to adult stem cells obtained from the adipose tissue of an individual. These adipose stem cells are multipotent stem cells capable of differentiating into various types of cells such as muscle cells, chondrocytes, and bone cells as well as adipocytes, and have the ability to multiply themselves. The adipocyte stem cells in the present invention are not limited thereto, but can be obtained mainly from human subcutaneous fat tissue by liposuction method, and preferably are autologous adipose stem cells obtained from their subcutaneous adipose tissue have.

In the above, the liposuction method is a method of collecting fat from the body using a syringe. In general, the method refers to subcutaneous fat tissue in the abdomen or buttock to inhale fat using a micro-catheter, and centrifuge the fat. Although a relatively large amount of cells can be extracted through such a method, cells not sufficiently differentiated into adipocytes can not be sure of differentiation into adipocytes related to hair growth after transplantation, and fully mature adipocytes can not be differentiated The efficiency is low, such as loss or deterioration of engraftment rate after transplantation, and in most cases, it is extracted in an amount insufficient for promoting hair growth or preventing / treating hair loss.

In the present invention, it is preferable that the minoxidil treated to increase the excipient activity of adipose stem cells is treated at a concentration of 1 to 1000 μM. When the adipose stem cell culture of the present invention is cultured in a medium supplemented with minoxidil for 24 to 96 hours, the enhancing ability can be enhanced. That is, the present invention can be carried out by inoculating adipose stem cells into a medium containing minoxidil and culturing for 24 to 96 hours.

Minoxidil is primarily an oral vasodilator and has been developed as an antihypertensive agent that directly lowers blood pressure by relaxing the smooth muscle of the peripheral artery. However, since hair growth effect is known as a external preparation, it is widely used as a treatment for male hair loss. In the present invention, it was confirmed that when minocystil was treated in adipocyte stem cell culture, the excitability of adipose stem cells was enhanced.

The present invention also provides a method for promoting the function of lipid stem cells, comprising culturing adipose stem cells in a medium containing minoxidil as an active ingredient.

The minoxidil contained in the culture medium of the present invention may be added at a concentration of 1 to 1000 μM as described above.

The culturing step of the present invention may be inoculation of adipose stem cells into a medium containing minoxidil and culturing for 24 to 96 hours.

In the present invention, cultivation can be carried out according to a suitable culture medium and culture conditions known in the art. Such a culturing process can be easily adjusted by those skilled in the art. These various methods of culturing are disclosed in various publications (e.g., James et al., Biochemical Engineering , Prentice-Hall International Editions). Suspension cultures and adherent cultures can be divided into batches, fed-batch and continuous-batch cultures according to the culture method.

The culture medium used for culturing should suitably satisfy the culture requirement of adipose stem cells. The medium includes various carbon sources, nitrogen sources, trace element components, growth factors, and the like. Examples of carbon sources that can be used include carbohydrates such as glucose, sucrose, lactose, fructose, maltose, starch and cellulose, fats such as soybean oil, sunflower oil, castor oil, coconut oil, palmitic acid, stearic acid, linoleic acid , Alcohols such as glycerol and ethanol, and organic acids such as acetic acid. These carbon sources may be used alone or in combination. Examples of nitrogen sources that can be used include organic nitrogen sources such as peptone, yeast extract, gravy, malt extract, corn steep liquor (CSL), and soybean wheat, and urea, such as ammonium sulfate, ammonium chloride, ammonium phosphate, ammonium carbonate and ammonium nitrate And an inorganic nitrogen source. These nitrogen sources may be used alone or in combination. The medium may include potassium dihydrogenphosphate, dipotassium hydrogenphosphate and the corresponding sodium-containing salts as a source. It may also include metal salts such as magnesium sulfate or iron sulfate. In addition, amino acids and suitable precursors and the like may be included. During the culture, compounds such as ammonium hydroxide, potassium hydroxide, ammonia, phosphoric acid and sulfuric acid can be added in a suitable manner to adjust the pH. In addition, foaming can be suppressed by using a defoaming agent such as fatty acid polyglycol ester during the culture. Preferably, the culture medium used in the present invention may be a medium commonly used in the field of animal cell culture, and commercially available DMEM (Dulbecco's Modified Eagle Medium), DMEM-F12 (mix of DMEM and Ham's F-12 medium) , Medium Essential Medium (MEM), Iscove's Modified Dulbecco's Medium (IMDM), MEM Alpha Modification, RPMI 1640 (Roswell Park Memorial Institute 1640), M199 / F12 (Medium 199 and Ham's F-12 medium) (Improved MEM), MCDB 131 medium and the like can be used. Minoxidil may be further added to the medium as described above. In addition, if necessary, it may further contain fetal bovine serum (FBS) or a growth factor or a differentiation-related factor, and may contain antibiotics conventionally used for stem cell culture, such as penicillin, Antibiotics such as streptomycin, kanamycin, ampicillin or amphotericin B may be added alone or in combination. At the time of culturing, CO ₂ can be maintained at a constant concentration (for example, 5%) as in the culture conditions of ordinary animal cells. The temperature at the time of culturing may be usually maintained at 20 캜 to 45 캜, preferably at 25 캜 to 40 캜.

After the completion of the culture, centrifugation or filtration may be carried out to separate the culture medium from the adipose stem cells, and these steps may be carried out as required by those skilled in the art.

In one embodiment of the present invention, migration of adipose stem cells and tube formation were observed after minoxidil treatment in order to confirm the activation of adipose stem cells by minoxidil (FIG. 1).

In another embodiment of the present invention, an increase in the expression of CXCL1 (chemokine (CXC motif) ligand 1) and PD-ECGF (platelet-derived endothelial cell growth factor) in order to examine the increase of paracrine effect of minoxidil- Respectively.

In another embodiment of the present invention, minoxidil-treated adipose stem cells were transplanted to the back region of the hair-removed mouse to confirm the hair-producing effect. The control group was compared with a group injected with only saline solution (vehicle) and a group implanted with adipose stem cells not activated with minoxidil. As a result, it was found that the group transplanted with minoxidil-activated adipose stem cells had faster and more abundant hair and had better hair growth effect than the control group. In particular, it was found that the activation of minoxidil results in a higher enhancing ability of adipose stem cells.

Accordingly, the present invention provides a composition for promoting hair loss prevention or hair growth comprising minoxidil-treated adipose stem cells.

The term " hair loss " of the present invention means a phenomenon in which hair completely escapes from the scalp. Minoxidil-treated adipose stem cells or cultures thereof, as demonstrated in the Examples below, convert hair from a resting phase to a growing phase, and the hair cells in the skin tissue It was confirmed that there is an effect of preventing, ameliorating, or treating hair loss, which increases the expression of growth factors associated with hair growth and wool.

The term " hair growth " of the present invention means that the hair grows in the scalp, and specifically refers to inducing hair follicles to form hair follicles at hair loss areas or hairless areas (hairless area). This is used synonymously with " hair growth " or " wool " which means that the length of hair is lengthened in the art (i.e., hair growth).

Specifically, the present invention provides a pharmaceutical composition for preventing, treating or promoting hair loss, which comprises minoxidil-treated adipose stem cells as an active ingredient.

The pharmaceutical composition according to the present invention may further contain, alone or in combination with one or more pharmaceutically acceptable carriers, excipients or diluents, the enhancer-enhancing adipose stem cells according to the method of the present invention. The term " pharmaceutically acceptable " as used herein refers to a composition that is physiologically acceptable and does not normally cause an allergic reaction or the like when administered to a human.

Pharmaceutically acceptable carriers may contain a variety of ingredients such as buffer, injectable sterile water, normal saline or phosphate buffered saline, sucrose, histidine, salts and polysorbates.

The pharmaceutical composition of the present invention can be administered orally or parenterally, and can be administered in the form of a general pharmaceutical preparation, for example, various forms of oral and parenteral administration at the time of clinical administration. In the case of formulation, , An extender, a binder, a wetting agent, a disintegrant, a surfactant, and the like.

Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may be prepared by mixing the pharmaceutical composition of the present invention with at least one excipient such as starch, calcium carbonate, Sucrose, lactose, gelatin, and the like.

In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included .

Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used as the non-aqueous solvent and suspension agent. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

In the present invention, when parenterally administered, the pharmaceutical composition may be administered by intravenous infusion, subcutaneous injection, muscle injection, intraperitoneal injection, transdermal administration or the like, and it is preferable that the administration route is determined depending on the kind of the applied disease Do. For example, the medicinal composition of the present invention is preferably used in a manner applied topically to the skin since it is used for prevention and treatment of hair growth stimulation or hair loss. Therefore, the pharmaceutical composition for promoting hair growth of the present invention is preferably an injection for topical administration, but is not limited thereto.

The pharmaceutical composition of the present invention can provide a desired alopecia prevention, treatment or hair growth promoting effect when an effective amount of an adipose stem cell or a culture thereof is contained. In the present invention, the term "effective amount" means the amount of the composition that exhibits the effect of preventing, treating or promoting hair loss. The effective amount of the adipose stem cells or culture thereof contained in the composition of the present invention will vary depending on the form in which the composition is commercialized, the method in which the compound is applied to the skin, and the time on the skin. For example, when the composition is commercialized as a medicament for alopecia prevention or treatment, it may contain adipocyte stem cells or a culture medium thereof at a higher concentration than a cosmetic product that is routinely applied to skin.

In addition, the appropriate dosage of the pharmaceutical composition or therapeutic agent of the present invention varies depending on factors such as the formulation method, administration method, age, body weight, sex, disease symptom, administration time, administration route and response sensitivity of the patient, Usually, a skilled physician can easily determine and prescribe dosages effective for the desired treatment. The minoxidil-treated adipose stem cells of the present invention have the same effect of promoting the same hair growth even when a small amount of cells are used in cell therapy, thereby reducing the dose of adipose stem cells in cell therapy.

The present invention also provides a quasi-drug composition for hair loss prevention or hair growth improvement comprising minoxidil-treated adipose stem cells as an active ingredient.

When the minoxidil-treated adipose stem cells of the present invention are used as an active ingredient of a quasi-drug composition, the adipose stem cells or the culture thereof exhibiting an effect of preventing hair loss or improving hair growth can be directly added, or can be used together with other quasi-drugs or quasi drugs. And can be appropriately used according to a conventional method. The amount of the active ingredient to be mixed can be appropriately determined depending on the purpose of use.

The quasi-drug composition for hair loss prevention or hair growth improvement of the present invention is not particularly limited in its formulation and may be variously formulated in the form of a quasi-drug known in the art to exhibit hair loss prevention or hair growth promoting effect. The formulated quasi-drug may be an ointment, a cream, a gel, a liquid, an emulsion, a patch or a spray, but is not limited thereto. Specifically, hair tonic, hair lotion, hair cream, hair spray, hair mousse, hair gel, hair conditioner, hair shampoo, hair rinse, hair pack, hair treatment, eyebrow hair extender, eyelash hair extender, eyelash nutrient, pet shampoo, pet Animal rinse, hand cleansing agent, detergent, soap, disinfectant cleaner, wet tissue, mask, ointment, patch or filter filler, and includes quasi-drugs in the conventional sense.

In addition, the quasi-drug composition for preventing hair loss or accelerating hair growth in each formulation can be blended with other components arbitrarily selected depending on the formulation of other quasi-drugs, the purpose of use, and the like. The amount of the active ingredient to be mixed may be appropriately determined depending on the purpose of use, and may include, for example, customary adjuvants such as thickeners, stabilizers, solubilizers, vitamins, pigments and flavors, and carriers.

In another aspect, the present invention provides a cosmetic composition for preventing hair loss or improving hair growth comprising minoxidil-treated adipose stem cells or a culture thereof as an active ingredient.

In the present invention, the cosmetics may be manufactured in any form conventionally produced in the art, and examples thereof include hair tonic, hair conditioner, hair essence, hair lotion, hair nutrition lotion, hair shampoo, hair rinse, Hair cosmetic cream, hair cream, hair cream, hair cream, hair cream, hair cream, hair cream, hair cosmetic cream, hair wax, hair aerosol, hair pack, hair nutrition pack, hair soap, hair cleansing foam, But the present invention is not limited thereto, and may be manufactured in various forms such as a waving agent for hair, a hair bleaching agent, a hair gel, a hair glaze, a hair dresser, a hair lacquer, a hair moistener, a hair mousse and a hair spray.

In the present invention, cosmetics may contain not only effective components such as minoxidil-treated adipocyte stem cells or cultures thereof, but also components commonly used in cosmetic compositions, such as antioxidants, stabilizers, solubilizers, vitamins, , ≪ / RTI > and carriers.

The term " promoting hair growth " or " hair loss prevention ", as used herein, is meant to include the use of other term wool or hair growth promoters as used in the art.

The part to which the hair growth stimulating medicinal composition or cosmetic of the present invention can be applied can be applied to any part of the body that requires hair growth as well as the scalp. For example, it can be used for areas with damaged hair or hairs due to trauma, or for improving the condition of wide forehead or M-type forehead, eyelashes or eyebrows and asphyxia for simple cosmetic effects.

In the present invention, a telogen to anagen transition model of C3H / HeN mice was used to compare the induction of the drug to anagen at about 7 weeks of age.

The term " anagen " of the present invention refers to the step of growing hair that accounts for 90% of the cycle of repeated growth of hair, and the term " telogen " It refers to the period when the cell division of the part of the hair part of the mother is stopped and the hair is temporarily stopped and the hair is pulled out.

Minoxidil-treated adipose stem cells according to the present invention showed remarkable increase in the proliferative capacity and migratory ability and increased secretion of growth factors. When administered to the animal model, the hair growth promoting effect and hair growth promoting effect were significantly increased as compared with the control group. Accordingly, the minoxidil-treated adipose stem cells according to the present invention are useful as a composition for preventing hair loss, treating hair loss or treating atopy, improving hair growth and promoting hair growth. Specifically, the present invention provides hair loss preventive hair growth It can be usefully used as an improvement drug composition, a quasi-drug composition or a cosmetic composition.

Brief Description of Drawings Figure 1 shows the effect of improving the migration (A, B) and tube forming ability (C) of adipose stem cells according to minoxidil treatment (A, Scratch analysis, B, Formation test results).
Fig. 2 shows the results of confirming an increase in expression of growth factors by minoxidil treatment.
FIG. 3 shows the result of confirming the increase in hair growth effect of adipose stem cells by minoxidil treatment (A, result of hair regeneration; B, result of hair regeneration regenerated on the 14th day of experiment).

Below, The present invention will be described in more detail by way of examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not construed as being limited by these embodiments.

Example 1. Adipose-derived stem cells from adipose tissue DerivedStemCell ) And culture

From subcutaneous fat, a known method (Kim et al. J Dermatol Sci 53: 96-102, 2009). Human fat-derived stem cells were obtained by liposuction. Specifically, the human adipose tissue supplied from the hospital was washed twice with PBS (a commonly used buffer), and then 0.075% collagenase was added thereto, followed by shaking for 50 minutes in a 37 ° C incubator. Subsequently, a culture medium in which a mixture of PBS and α-MEM medium was added to the above culture medium was centrifuged at 1200 rpm, and the supernatant was removed and only the material pelleted into the pellet was used. PBS was added to the separated pellet, and the mixture was vigorously mixed. Then, the mixture was filtered through a 100 μm nylon mesh, and centrifuged to obtain pellets. The obtained fat stem cells, 10% FBS (GIBCO, Invitrogen) , 1% penicillin / streptomycin α-MEM medium supplemented with (GIBCO) containing (Hyclone, Thermo Scientific), and cultured at 37 ℃, 5% CO ₂ to 7 ~ 9 passages were used. The positive expression of CD44, CD73, CD90, CD105, HLA-1 and PODXL and negative expression of CD34 and CD45 were confirmed by flow cytometry analysis (Kim et al. Cell Death Dis 　 4: e 588, 2013; Kim et al., Cell Biol Int 38: 32-40, 2014), confirming the multipotent ability to differentiate into adipocytes, bone cells and chondrocytes (Song et al., Stem Cells Dev 　 17: 451-461, 2008).

Example  2. Minoxidil  Migration and tubing of adipose stem cells by treatment Formability  Improvement effect

In order to confirm the improvement of migration ability of adipose stem cells according to minoxidil treatment, a scratch assay and a transfer ability test using a transwell were performed.

Specifically, for the scratch analysis, the same number of cells were cultured until completely filled in a 6 well dish, and then scratches were generated using appropriate tools. FBS-free medium and 20 μM, 50 μM minoxidil The media were separated by the addition of media and observed for a certain period of time. Photographs were taken and analyzed. As a result, as shown in Fig. 2A, it was found that the number of cell migrations increased after treatment of adipose stem cells with minoxidil.

In addition, in the test of the ability to move using a transwell, the same number of cells were seeded in a transwell. Then, in a medium containing no FBS and 20 μM of minoxidil at a concentration of 50 μM, And then stained with a crystal dye to analyze the number of cells transferred through the transwell membrane. As a result, as shown in Fig. 2B, it was found that the migration ability of adipose stem cells was improved in a concentration-dependent manner after minoxidil treatment.

In order to confirm the increase in tube formability of adipose stem cells following minoxidil treatment, matrigel was coated on a 12-well dish, and the same number of cells were seeded and EBM endothelial basal cell medium) and minoxidil were added and incubated for a certain period of time. Then, pictures were taken and the number of tubes was analyzed. As a result, as shown in Fig. 2C, it was found that the tube forming ability of adipose stem cells was increased depending on the concentration of minoxidil.

Example  3. Minoxidil  Treatment of adipose stem cells Paracrine  Increase effect

Specifically, adipose stem cells were treated with 20 μM and 50 μM of minoxidil, and after 3 days, the membranes were broken with Trizol. Next, 1/5 volume of chloroform of the whole sample was added, and the mixture was vigorously mixed and centrifuged at 14000 rpm for 15 minutes. The supernatant was carefully collected, mixed with the same amount of isopropanol, kept frozen for 30 minutes or more, and then centrifuged at 14000 rpm for 15 minutes. To obtain only pellet, the supernatant was discarded, washed once with 70% ethanol, and the obtained pellet was dried to isolate RNA. CDNA was synthesized by reacting the identified RNA with reverse transcriptase. Real-time PCR was performed by adding the generated cDNA, a suitable primer, and a SYBR enzyme mixture, and then corrected by GAPDH, a house keeping gene.

The primer sequences used in this Example are shown in Table 1 below.

designation order SEQ ID NO: GAPDH forward 5'-CGAGATCCCTCCAAAATCAA-3 ' One GAPDH reverse 5'-TGTGGTCATGAGTCCTTCCA-3 ' 2 CXCL1forward 5'-CATCCAAAGTGTGAACGTGA-3 ' 3 CXCL1 reverse 5'-CTTAACTATGGGGGATGCAG-3 ' 4 PD-ECGF forward 5'-GCTGTATCGTGGGTCAGAGT-3 ' 5 PD-ECGF reverse 5'-AGAATGGAGGCTGTGATGAG-3 ' 6

As a result, as shown in Fig. 2, the expression of CXCL1 and PD-ECGF, which are known to be related to the hair growth of the cell growth factors after minoxidil treatment, were significantly increased.

Example  4. Minoxidil  Treatment of adipose stem cells Hair growth effect  increase

In the present Example, it was confirmed that the hair growth effect was increased by treating mincedilide in adipose stem cells.

Specifically, hair of C3H / HeN mice at 7 weeks of age, which is a telogen to anagen transition model in the resting phase of the hair cycle, was shaved and treated with FBS-treated 1 x 10 ⁴ adipose stem cells (control) and 10 (Kim et al., Stem Cells Dev 23: 1364-1376, 2014) were injected subcutaneously into the mice, respectively, in the adipocyte stem cells (experimental group) pretreated with ng / ml minoxidil for 24 hours. Skin darkening, a sign that the hair cycle was induced, was monitored and weight was measured after shaving the dorsal hair 15 days later.

As a result, the growth induction of hair was not limited to the site of cell injection, and skin darkening or hair regeneration appeared at all sites such as (Fig. 3A). In addition, the weight of the hair measured 14 days later was significantly increased more than three times in the fat stem cell group pretreated with minoxidil (FIG. 3B).

<110> Industry-Academic Cooperation Foundation, Yonsei University <120> Composition for enhancing the action for hair growth of adipose          Minoxidil as an active ingredient <130> 1063041 <160> 6 <170> KoPatentin 3.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> GAPDH forward <400> 1 cgagatccct ccaaaatcaa 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> GAPDH reverse <400> 2 tgtggtcatg agtccttcca 20 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CXCL1forward <400> 3 catccaaagt gtgaacgtga 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> CXCL1 reverse <400> 4 cttaactatg ggggatgcag 20 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> PD-ECGF forward <400> 5 gctgtatcgt gggtcagagt 20 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> PD-ECGF reverse <400> 6 agaatggagg ctgtgatgag 20

Claims (8)

A composition for promoting the function of lipid stem cells comprising Minoxidil as an active ingredient.
The method according to claim 1,
Wherein the minoxidil is contained at a concentration of 1 to 1000 μM.
A method for promoting the promoting activity of adipose stem cells comprising culturing adipose stem cells in a medium containing minoxidil as an active ingredient.
The method of claim 3,
Wherein the minoxidil is added at a concentration of 1 to 1000 μM.
The method of claim 3,
Wherein the culturing is performed by adding minoxidil to an adipocyte stem cell culture medium and culturing for 24 hours to 96 hours.
The method of claim 3,
DMEM-F12 (mix of DMEM and Ham's F-12 medium), MEM (Minimum Essential Medium), IMD (Iscove's Modified Dulbecco's Medium), α-MEM (MEM Alpha Modification) Wherein the medium is at least one selected from the group consisting of RPMI 1640 (Roswell Park Memorial Institute 1640), M199 / F12 (Medium 199 and Ham's F-12 medium), IMEM (Improved MEM) and MCDB 131 medium Enhancing method.
A pharmaceutical composition for promoting prevention, treatment or hair growth of alopecia comprising an adipocyte stem cell treated with minoxidil by the method of any one of claims 3 to 6 or a culture thereof as an active ingredient.
7. A cosmetic composition for preventing hair loss or improving hair growth, which contains minoxidil-treated adipose stem cells or a culture thereof as an active ingredient by the method according to any one of claims 3 to 6.

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