WO2014178438A1 - Préparation de cellules pour la régénération capillaire - Google Patents

Préparation de cellules pour la régénération capillaire Download PDF

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Publication number
WO2014178438A1
WO2014178438A1 PCT/JP2014/062164 JP2014062164W WO2014178438A1 WO 2014178438 A1 WO2014178438 A1 WO 2014178438A1 JP 2014062164 W JP2014062164 W JP 2014062164W WO 2014178438 A1 WO2014178438 A1 WO 2014178438A1
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Prior art keywords
adipose tissue
stem cells
cell preparation
mesenchymal stem
derived mesenchymal
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PCT/JP2014/062164
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English (en)
Japanese (ja)
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正典 佐伯
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Saeki Masanori
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Priority to JP2015514878A priority Critical patent/JP6362588B2/ja
Publication of WO2014178438A1 publication Critical patent/WO2014178438A1/fr

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    • 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
    • C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
    • C12N5/0662—Stem cells
    • C12N5/0667—Adipose-derived stem cells [ADSC]; Adipose stromal stem cells
    • 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
    • A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
    • A61K8/981—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of mammals or bird
    • 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
    • A61P17/14—Drugs for dermatological disorders for baldness or alopecia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q7/00—Preparations for affecting hair growth
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
    • A61K2800/86—Products or compounds obtained by genetic engineering
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations

Definitions

  • the present invention relates to a cell preparation for hair regeneration. More specifically, the present invention relates to a cell preparation for regenerating hair by regenerative medicine.
  • hair restorers containing minoxidil, capronium chloride, trans-3,4'-dimethyl-3-hydroxyflavanone, nicotinamide, piroctone olamine, adenosine and the like have been put into practical use.
  • the conventional hair restorer only promotes the proliferation and activation of hair papilla cells, and the effect is said to be greatly limited by individual differences and is limited.
  • An object of the present invention is to provide a cell preparation capable of reproducing hair using stem cells in order to realize hair regeneration medicine using stem cells.
  • the present inventor made an intensive study to solve the above problems, and made the adipose tissue-derived mesenchymal stem cells coexist with adipocytes, and transplanted it to a site where hair growth is desired. It was found that hair growth was observed at the site, and the hair could be regenerated.
  • the present invention has been completed by further study based on this finding.
  • the present invention provides a cell preparation for hair regeneration having the following aspects: Item 1. A cell preparation for hair regeneration, comprising adipose tissue-derived mesenchymal stem cells and adipocytes. Item 2. Item 2. The cell preparation for hair regeneration according to Item 1, wherein 1 to 10 adipose tissue-derived mesenchymal stem cells are contained per one adipocyte. Item 3. Item 3. The cell preparation for hair regeneration according to Item 1 or 2, wherein the number of adipose tissue-derived mesenchymal stem cells contained per mL of cell preparation is 10,000 or more. Item 4. Item 4. The cell preparation for hair regeneration according to any one of Items 1 to 3, which is used for autotransplantation. Item 5. Item 5. Item 5.
  • the cell preparation for hair regeneration according to any one of Items 1 to 4, which is a concentrate of adipose tissue-derived mesenchymal stem cells and adipocytes obtained by removing a liquid fraction from the collected adipose tissue .
  • Items 1 to 4 which is a concentrate of adipose tissue-derived mesenchymal stem cells and adipocytes obtained by removing a liquid fraction from the collected adipose tissue .
  • Item 6 Use of a cell preparation containing adipose tissue-derived mesenchymal stem cells and adipocytes for the production of a hair regenerating agent.
  • Item 7. A method of regenerating hair, comprising a step of administering a cell preparation containing adipose tissue-derived mesenchymal stem cells and adipocytes to a site where hair regeneration is required.
  • a cell preparation containing adipose tissue-derived ⁇ mesenchymal stem cell (AD-MSC, AT-MSC) and adipocytes is transplanted to a site where hair growth is desired.
  • AD-MSC adipose tissue-derived ⁇ mesenchymal stem cell
  • AT-MSC AT-MSC
  • adipocytes adipose tissue-derived ⁇ mesenchymal stem cell
  • adipocytes adipose tissue-derived ⁇ mesenchymal stem cell
  • the present invention also has the advantage that the cell preparation can be used as it is for transplantation, and it does not require tissue transplantation once hair follicles have been formed, so that the hair can be regenerated with almost no trace left at the transplant site.
  • pigment cells melanocytes
  • hair matrix cells since pigment cells (melanocytes) can be induced together with the hair matrix cells, it is possible to regenerate black natural hair instead of white hair.
  • the cell preparation of the present invention is used for hair regeneration and is characterized by containing adipose tissue-derived mesenchymal stem cells and adipocytes.
  • adipose tissue-derived mesenchymal stem cells and adipocytes.
  • Adipose tissue-derived mesenchymal stem cells are stem cells that can differentiate into adipose tissue, muscle cells, bones, cartilage, ligaments, skin cells, and nerve cells.
  • the adipose tissue-derived mesenchymal stem cells used in the present invention may be collected from living adipose tissue, or cultured adipose tissue-derived mesenchymal stem cells collected from a living adipose tissue It may be.
  • the adipose tissue-derived mesenchymal stem cells used in the present invention are preferably those collected from a living body from the viewpoint of suppression of immune reaction.
  • the adipose tissue-derived mesenchymal stem cells present in the subcutaneous adipose tissue can be easily collected with relatively low attack, can be collected in a relatively large amount, and does not require growth by culture, and can efficiently regenerate hair.
  • the adipose tissue-derived mesenchymal stem cells present in the subcutaneous adipose tissue can be easily collected with relatively low attack, can be collected in a relatively large amount, and does not require growth by culture, and can efficiently regenerate hair.
  • it can be collected at the same time as fat cells, and it is preferably used in the present invention.
  • the method for obtaining the adipose tissue-derived mesenchymal stem cells is not particularly limited, and preferably includes the method described in the section “Method for preparing cell preparation” described later.
  • a fat cell is a cell that has the function of synthesizing, storing, and releasing fat, and that forms adipose tissue.
  • Adipocytes do not undergo cell division because they are completely differentiated.
  • the fat cells include white fat cells and brown fat cells. In the present invention, any of these may be used, or a mixture of both may be used.
  • the adipocytes used in the present invention may be used in the state in which adipocytes are isolated, or by using a mixture of extracellular matrix (ECM) and adipocytes present around the adipocytes. Also good.
  • ECM extracellular matrix
  • the collected adipose tissue may be used as it is as the adipocyte, or one obtained by removing the extracellular matrix from the collected adipose tissue may be used.
  • the concentration of the adipose tissue-derived mesenchymal stem cells in the cell preparation of the present invention is not particularly limited, but for example, from the viewpoint of efficiently promoting hair regeneration, for example, the adipose tissue-derived mesenchyme contained per mL of cell preparation
  • the number of stem cells is 10,000 or more, preferably 100,000 or more, more preferably 150,000 or more, and particularly preferably 150,000 to 200,000.
  • the concentration of adipocytes in the cell preparation of the present invention is not particularly limited, but from the viewpoint of efficiently promoting hair regeneration, for example, the number of adipocytes contained in 1 mL of cell preparation is 1,000.
  • the number is preferably 5,000 or more, more preferably 10,000 or more, and particularly preferably 10,000 to 20,000.
  • the ratio between the adipose tissue-derived mesenchymal stem cells and the adipocytes is not particularly limited, and a more excellent hair regeneration effect is obtained as the content of the adipose tissue-derived mesenchymal stem cells is increased.
  • at least one adipose tissue-derived mesenchymal stem cell per adipocyte preferably 5 or more, more preferably 1 to 10, more preferably 5 to 10 Can be mentioned.
  • the cell preparation of the present invention immune rejection can be avoided by using patient-derived adipocytes and adipose tissue-derived mesenchymal stem cells to be administered for the purpose of autologous transplantation.
  • the cell preparation of the present invention is prepared using adipocytes derived from a person other than the patient to be administered and / or adipose tissue-derived mesenchymal stem cells, and is not limited to use in other transplants.
  • adipocytes extracellular matrix (ECM) present around the adipocytes) collected from others before transplantation are used.
  • ECM extracellular matrix
  • adipose tissue-derived mesenchymal stem cells are usually cells that do not have immunogenicity, but those subjected to the treatment may be used as necessary.
  • the collected adipose tissue may be used as it is as the cell preparation of the present invention.
  • Removed impurities from the adipose tissue for example, anesthetic fluid (so-called tumecent solution), aged adipocytes, blood, tissue fluid, etc. injected during the collection of fat), impurities and some of the collected adipose tissue You may use what removed the fat cell.
  • the method for preparing the cell preparation include methods (a) to (c) described later.
  • ⁇ Other ingredients> conventionally known pharmaceutically acceptable carriers, excipients, anti-inflammatory agents, analgesics, immunosuppressants, and the like can be added to the cell preparation of the present invention as necessary. Furthermore, collagen, fibroblasts, growth factors, growth factors, cytokines, etc. may be added as long as the effects of the present invention are not impaired.
  • TGF ⁇ 1 transforming growth factor ⁇ 1
  • the cell preparation of the present invention may contain cells such as adipose stem cells.
  • the cell preparation of the present invention comprises diffuse alopecia, postpartum alopecia, androgenetic alopecia, seborrheic alopecia, senile alopecia, alopecia areata, scarring alopecia, hair loss caused by mental disorders, etc. Can improve thinning hair.
  • the dosage of the cell preparation of the present invention is appropriately set according to the amount of hair at the administration site, the patient's sex, physique, etc. For example, 1,000 cm in terms of the number of fat cells per cm 2 of the administration site. Up to about 100,000, preferably about 1,000 to 50,000, and more preferably about 5,000 to 10,000. Further, for example, when the cell preparation obtained by the following method (c) is used, for example, 0.1 to 1.0 mL, preferably 0.1 to 0.5 mL, more preferably 0 per 1 cm 2 of the administration site. .2 to 0.5 mL.
  • the method for administering the cell preparation of the present invention to a site where hair regeneration is required is not particularly limited, and a conventionally known administration method can be adopted, for example, a method of injecting with a syringe, cannula or the like. It is done.
  • the hair regeneration effect of the cell preparation of the present invention varies among individuals, but usually hair growth is observed about 2 to 6 weeks after administration, and the hair is regenerated at the site.
  • the cell preparation of the present invention may be obtained by collecting and mixing adipose tissue-derived mesenchymal stem cells and adipocytes, or collected from adipose tissue as a mixture of adipose tissue-derived mesenchymal stem cells and adipocytes. You may use what you did. In particular, it is preferable to obtain a concentrate of adipose tissue-derived mesenchymal stem cells and adipocytes by removing a liquid fraction (such as a body fluid) from the adipose tissue to obtain the cell preparation of the present invention. More simply, the adipose tissue collected as the cell preparation of the present invention may be used as it is.
  • preferred methods include the following methods (a) to (c). According to the methods (a) to (c), a liquid fraction (body fluid or the like) can be removed from the adipose tissue to obtain a concentrate of adipose tissue-derived mesenchymal stem cells and adipocytes.
  • a liquid fraction body fluid or the like
  • Method (a) for preparing the cell preparation of the present invention includes the step (1a) of removing a liquid fraction from the collected adipose tissue to obtain a cell fraction.
  • the collected fat refers to a mixture of a cell fraction and a liquid fraction derived from adipose tissue collected from adipose tissue by liposuction, lipotomy, degreasing, and the like.
  • the cell fraction contains adipocytes, adipose stem cells, and impurities (blood cells, dead / alive cells, senescent cells, etc.).
  • the liquid fraction includes a tumecent liquid, a cell tissue liquid, and the like.
  • the separation of the liquid fraction and the cell fraction can be carried out simply by allowing the collected fat to stand and allowing the cell fraction to settle.
  • the method for removing the liquid fraction is not particularly limited, and examples thereof include decantation and suction. Moreover, you may perform centrifugation, filtration, etc. as needed.
  • Centrifugation and filtration conditions are not particularly limited as long as they do not damage adipocytes and adipose tissue-derived mesenchymal stem cells and can remove impurities, but for example, 700 to 2500 ⁇ g for 1 to 15 minutes, preferably 2000 to Examples include conditions at 2200 ⁇ g for 5 to 10 minutes.
  • the collected fat is subjected to a mesh filter having a hole size of, for example, 10 to 300 ⁇ m, preferably 10 to 150 ⁇ m, more preferably 10 to 100 ⁇ m, and still more preferably 15 to 20 ⁇ m.
  • the mixture of adipocytes and adipose tissue-derived mesenchymal stem cells can be obtained on the filter by removing blood, tissue fluid, anesthetic agent (tumecent solution) and the like administered to the site where the fat is collected.
  • a container including such a mesh filter may be filled with collected fat containing impurities and subjected to centrifugation.
  • adipose tissue containing adipose tissue-derived mesenchymal stem cells and adipocytes (including extracellular matrix) from which the liquid fraction has been removed can be obtained, and this can be used as the cell preparation of the present invention. Can do.
  • the method (b) for preparing the cell preparation of the present invention includes the following steps (1b) to (3b). (1b) removing the liquid fraction from the collected adipose tissue to obtain a cell fraction; (2b) a step of separating adipose tissue-derived mesenchymal stem cells and adipocytes from at least a part of the cell fraction obtained in the step (1b), and (3b) a cell fraction obtained in the step (1b). A step of mixing a part of a minute or a part of the adipocyte obtained in the step (2b) and the adipose tissue-derived mesenchymal stem cell obtained in the step (2b).
  • step (1b) is as described in the above method (1a).
  • step (2b) adipose tissue-derived mesenchymal stem cells are separated from at least a part, preferably half, of the cell fraction obtained in the step (1b).
  • the adipose tissue-derived mesenchymal stem cells obtained in the step (2b) are used as a part of the cell fraction obtained in the step (1b) or the fat obtained in the step (2b).
  • a cell preparation containing adipose tissue-derived mesenchymal stem cells and adipocytes can be obtained.
  • examples of the method for separating adipose tissue-derived mesenchymal stem cells include the method described in JP-T-2005-519883. More specifically, a proteolytic enzyme is added to the cell fraction in order to facilitate separation of adipose tissue-derived mesenchymal stem cells and adipocytes by decomposing bonds between cells.
  • proteolytic enzymes include collagenase, trypsin, lipase, etc., and one kind selected from these may be used alone, or two or more kinds may be used in combination.
  • the method for separating adipose tissue-derived mesenchymal stem cells and adipocytes from the enzyme-treated cell fraction is not particularly limited and may be appropriately selected from conventionally known methods. Examples thereof include centrifugation and filtration. .
  • the centrifugation conditions are not particularly limited as long as they can be separated without damaging the adipose tissue-derived mesenchymal stem cells and adipocytes.
  • the conditions are 100 to 500 ⁇ g for 1 to 20 minutes, preferably 100 to 300. A condition of x to 5 to 10 minutes is mentioned.
  • the cell fraction or suspension thereof is passed through a mesh filter having a hole size of 10 to 100 ⁇ m, preferably 10 to 50 ⁇ m, more preferably 15 to 20 ⁇ m.
  • Adipose tissue-derived mesenchymal stem cells can be obtained on top, and adipocytes can be obtained in the filtrate.
  • the enzyme-treated adipose tissue-derived mesenchymal stem cells and adipocytes are preferably subjected to a washing treatment after separation, and washing is performed with a phosphate buffer solution (PBS), physiological saline, Ringer's solution, dextran, or the like. Can do.
  • PBS phosphate buffer solution
  • the washing treatment may be repeated a plurality of times as necessary, and may be subjected to centrifugation as necessary when cells are collected after washing.
  • the conditions for the centrifugation may be the same as those employed when separating the adipose tissue-derived mesenchymal stem cells and adipocytes from the enzyme-treated cell fraction.
  • the cell preparation thus prepared contains a high percentage of adipose tissue-derived mesenchymal stem cells together with adipocytes, and can exhibit an excellent hair regeneration effect.
  • the method (c) for preparing the cell preparation of the present invention comprises the following steps (1c) and (2c). (1c) removing the liquid fraction from the collected adipose tissue to obtain a cell fraction; and (2c) removing impurities from the cell fraction obtained in the step (1c) to obtain a mesenchymal system derived from adipose tissue Obtaining a concentrate of stem cells and adipocytes.
  • step (1c) is as described in the above method (1a).
  • impurities are removed from the cell fraction and concentrated.
  • the method for removing and concentrating impurities is not particularly limited and may be appropriately selected from conventionally known methods.
  • a syringe equipped with a weight filter described in JP-T-2007-533396 is used for convenience.
  • impurities such as free oil, dead and active cells, and senescent cells released from broken fat cells are removed, and a healthy adipose tissue-derived mesenchymal stem cell and fat cell concentrate can be obtained.
  • Such a syringe is commercially available. Specifically, a syringe manufactured for LIPOMAX-SC (manufactured by Medikan Corp., Seoul, Korea) is exemplified.
  • the collected fat is filled in a syringe equipped with a filter inside, and the fat is separated into a liquid fraction and a cell fraction by centrifugation, and free oil (drained oil) is further separated by a filter. ) Are separated.
  • the liquid fraction separated here is not completely separated in the step (1c) but is mixed in the cell fraction.
  • a concentrate of mesenchymal stem cells and adipocytes can be obtained, which can be used as the cell preparation of the present invention. Since the cell preparation obtained in this way has selected and concentrated healthy adipose tissue-derived mesenchymal stem cells and adipocytes, it is difficult to cause necrosis, calcification, etc. after transplantation, and diseases of the interosseous junction It is more useful for alleviating and treating the symptoms, and for repairing damaged muscles and alleviating pain due to muscle damage.
  • Centrifugation conditions for separating the fat into a liquid fraction and a cell fraction include impurities, adipose tissue-derived mesenchymal stem cells, and fat without damaging the adipose tissue-derived mesenchymal stem cells and fat cells.
  • the conditions are not particularly limited as long as the cells can be separated, but examples include conditions of 700 to 2500 ⁇ g for 1 to 15 minutes, preferably 2000 to 2200 ⁇ g for 5 to 10 minutes.
  • the pore size of the filter is not particularly limited as long as it does not allow adipose tissue-derived mesenchymal stem cells and adipocytes to pass through and allows impurities to pass through.
  • the pore size is 10 to 300 ⁇ m, preferably 10 to 150 ⁇ m.
  • the thickness is preferably 10 to 100 ⁇ m, more preferably 15 to 20 ⁇ m.
  • this method (c) separates adipose tissue-derived mesenchymal stem cells and adipocytes from a part of the concentrate obtained in step (2c), and further obtains the obtained adipose tissue-derived mesenchymal stem cells.
  • the method for separating the adipose tissue-derived mesenchymal stem cells is as described for the step (2b) of the method (b).
  • this step (3c ′) when adipose tissue-derived mesenchymal stem cells are separated using a proteolytic enzyme, usually about 700,000 or more, preferably about 1.5 million per cc of the concentrate obtained in step (2c).
  • One or more, more preferably about 2 million or more adipose tissue-derived mesenchymal stem cells can be obtained.
  • the cell preparation prepared through this step (3c ′) contains not only healthy adipocytes, but also a high concentration of adipose tissue-derived mesenchymal stem cells. There is an effect.
  • the fat cells in the obtained concentrate are refined and separated as a mixture of the adipose tissue-derived mesenchymal stem cells and the refined adipocytes.
  • (3c ′′) may be included.
  • Adipocytes can be miniaturized by a method that is usually employed in the art, and is not particularly limited. For example, adipose tissue-derived mesenchymal stem cells are cut after drilling fat cells with a drill or the like as necessary. And separating the micronized adipocyte mixture. As the separation method, the above-mentioned centrifugation conditions can be employed.
  • a cell preparation containing high-concentrated adipocytes together with adipose tissue-derived mesenchymal stem cells can be prepared.
  • the fine fat cells refer to fat cells having a size that can pass through an injection needle of 26 to 30 gauge, preferably 18 to 30 gauge.
  • the treatment for refining fat cells can be performed at least once, preferably 1 to several times, more preferably 1 to 2 times.
  • the adipose tissue-derived mesenchymal stem cells were separated from a part of the mixture of the refined adipose tissue-derived mesenchymal stem cells obtained in the step (3c ′′) and the refined adipocytes, and obtained.
  • the adipose tissue-derived mesenchymal stem cells are the concentrate obtained in the step (2c) or the refined adipose tissue-derived mesenchymal stem cells obtained in the step (3c '') and the refined adipocytes. (4c ′′) may be included in a part of the mixture.
  • the method for separating adipose stem cells is as described for step (2b) of method (b) above.
  • a syringe described in JP-T-2007-533396 can be equipped with a cannula or the like, filled with fat collected by liposuction, and directly subjected to removal of impurities, concentration treatment, and the like to obtain a cell preparation. . After that, when administering the cell preparation, it can be transferred directly from the syringe to the syringe for injecting the cell preparation and administered to the affected area, so that the cell preparation should be highly safe without problems such as contamination. it can.
  • this method (c) can also be carried out using a commercially available kit. Specific examples of such a kit include LIPOMAX-SC (manufactured by MEDIKAN Corp.). Is done.
  • the cell preparation of the present invention can be obtained by any of the above methods, but is preferably a cell preparation obtained by the method (b) or (c), more preferably a cell preparation obtained by the method (c), Preferably, the cell preparation obtained by the method (c) including the step (3c ′′), particularly preferably the cell preparation obtained by the method (c) including the step (3c ′′) and the step (4c ′′). .
  • the cell preparation obtained through the step (3c ′′) or the method (c) including the step (3c ′′) and the step (4c ′′) is finely mixed with adipose tissue-derived mesenchymal stem cells. It contains high-concentrated healthy fat cells and can exhibit a remarkable hair regeneration effect.
  • Liposuction was performed from each subject using a syringe (syringe manufactured for LIPOMAX-SC: manufactured by Medikan Corp., Seoul, Korea).
  • the collected fat-containing syringe was allowed to stand at room temperature (about 25 ° C.) for about 10 minutes to separate the cell fraction and the liquid fraction, and the liquid fraction containing the tumecent liquid was discarded.
  • the syringe was then centrifuged at 2200 ⁇ g for 8 minutes. The mixture was separated into three layers by centrifugation, an upper layer (free oil), an intermediate layer (adipocytes and adipose tissue-derived mesenchymal stem cells), and a lower layer (liquid fraction such as cell tissue fluid).
  • the fat cells and adipose stem cells derived from each patient were used, the above operation was performed for each patient.
  • the number of adipose tissue-derived mesenchymal stem cells and adipocytes in the obtained cell preparation varies depending on the patient, but about 1 to 200,000 adipose tissue-derived mesenchymal stem cells per ml of cell preparation and about adipocytes are present. 10,000 to 20,000 were included.

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Abstract

Le but de la présente invention est de fournir une préparation de cellules qui permet la régénération capillaire par utilisation de cellules souches, afin d'assurer la réalisation d'une thérapie de régénération capillaire par utilisation de cellules souches. Lorsque des cellules souches mésenchymateuses dérivées de tissu adipeux, qui sont dans un état de coexistence avec des adipocytes, sont transplantées dans une zone où la croissance de nouveaux cheveux est souhaitée, la croissance des cheveux est observée dans la zone et, par conséquent, les cheveux peuvent être régénérés.
PCT/JP2014/062164 2013-05-02 2014-05-02 Préparation de cellules pour la régénération capillaire WO2014178438A1 (fr)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019221307A1 (fr) * 2018-05-16 2019-11-21 地方独立行政法人神奈川県立産業技術総合研究所 Méthode de contrôle de la couleur de poils régénérés, méthode de régénération des poils, et méthode de production d'ébauches de follicules pileux
WO2020009147A1 (fr) 2018-07-04 2020-01-09 正典 佐伯 Formulation de filtrat de cellules souches et son procédé de préparation
CN113750117A (zh) * 2021-09-30 2021-12-07 四川大学 从吸脂废液中制备的脂肪组织精华、其制备方法和应用
EP4151719A1 (fr) * 2015-12-30 2023-03-22 Alfredo Gorio Procédé pour favoriser et améliorer les propriétés du tissu adipeux, tissu et cellules obtenus par ledit procédé

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