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WO2012110232A1 - Procédé de production d'une préparation en utilisant des cellules du follicule pileux, préparation et kit - Google Patents

Procédé de production d'une préparation en utilisant des cellules du follicule pileux, préparation et kit Download PDF

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Publication number
WO2012110232A1
WO2012110232A1 PCT/EP2012/000657 EP2012000657W WO2012110232A1 WO 2012110232 A1 WO2012110232 A1 WO 2012110232A1 EP 2012000657 W EP2012000657 W EP 2012000657W WO 2012110232 A1 WO2012110232 A1 WO 2012110232A1
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WO
WIPO (PCT)
Prior art keywords
cells
buffer solution
kit
hair follicles
buffer
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/EP2012/000657
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German (de)
English (en)
Inventor
Wolfgang Richter
Christian Toloczyki
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
EURODERM GmbH
Original Assignee
EURODERM GmbH
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from DE102011011623A external-priority patent/DE102011011623A1/de
Application filed by EURODERM GmbH filed Critical EURODERM GmbH
Priority to EP12705789.1A priority Critical patent/EP2768947A1/fr
Publication of WO2012110232A1 publication Critical patent/WO2012110232A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • 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
    • C12N5/0625—Epidermal cells, skin cells; Cells of the oral mucosa
    • C12N5/0627—Hair cells
    • 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
    • C12N2509/00—Methods for the dissociation of cells, e.g. specific use of enzymes

Definitions

  • the present invention relates to a method for producing a preparation according to claim 1. It further relates to a preparation according to claim 22 and a kit according to claim 23.
  • body cells such as the neural crest, derived cells, e.g. Melanocyte progenitor cells, and / or keratinocyte progenitor cells, for example, for increasing or enhancing pigmentation of certain areas of the skin, are known, i.a. from PCT / EP 2008/007684 assigned to the assignee of the present invention, the content of which is hereby incorporated in its entirety by reference into the present application.
  • PCT / EP 2008/007684 also discloses a method for preparing the body cells used.
  • An object of the present invention is to provide another method for producing a preparation having cells of the body or for preparing the preparation or the cells.
  • This object of the present invention is achieved by a method according to claim 1.
  • a preparation with cells according to claim 22 and a kit for producing a preparation according to claim 23 are proposed.
  • a method for producing a preparation in particular a suspension comprising cells of the human body is proposed.
  • the separation of the buffer solution by filtration to give a last filter residue is an optional step that is not subject matter
  • CONFIRMATION COPY certain embodiments.
  • a continuous washing process is provided in which the enzyme solution gradually dilutes out.
  • no such washing process or separation is provided.
  • the preparation according to the invention which was produced using the method according to the invention, has the features of claim 22.
  • a kit is proposed with which the method according to the invention can be carried out and by means of which the preparation according to the invention can be produced.
  • the method of the invention does not include applying a preparation or product produced therewith to a body portion of a patient and / or injecting the produced preparation or product into a body portion and / or other forms of application.
  • the cells used are, for example, melanocyte progenitor cells, and / or keratinocyte progenitor cells and / or mesenchymal progenitor cells (hereinafter also referred to as "progenitor cells") and / or neural crest-derived cells, or in association or in a group or
  • progenitor cells are also present in a cell mixture which is obtained if the cells obtained from the hair root sheath are not separated according to their origin, function or nature, but if the cells obtained are separated on receipt, they can pass through Use of primarily neural crest-derived cells, eg, melanocyte progenitor cells, and / or keratinocyte progenitor cells and / or mesenchymal progenitor cells, to achieve a higher effect after their application in accordance with the present invention or make it more suitable for a specific application.
  • neural crest cells e.g. Melanocyte progenitor cells or stem cells and / or
  • Keratinocyte precursor cells or stem cells this also applies without restriction to mesenchymal precursor cells and stem cells, even if this is not expressly mentioned.
  • PBS Phosphate Buffered Saline, phosphate-buffered saline
  • biocompatible, viscosity-enhancing substances such as e.g. Hyaluronan or collagen.
  • cells originating from the neural crest are understood as meaning both autologous and allogeneic and xenogenic cells originating from the neural crest, for example melanocyte progenitor cells or stem cells
  • melanocyte progenitor cells include both autologous and allogeneic and xenogenic eratinocyte progenitor cells or stem cells
  • melanocyte progenitor cells include both autologous and allogeneic and xenogenic eratinocyte progenitor cells or stem cells
  • mesenchymal progenitor cells include both autologous and allogeneic and xenogeneic mesenchymal precursor cells or stem cells.
  • the cells of the invention have been obtained by plucking hair from the vital skin of the donor or by plucking or otherwise harvesting hair from a present skin biopsy of the donor.
  • the cells are recovered and intended for use without first being spent or spent in a culture to grow cells.
  • the cells are not or will not be altered, particularly not genetically altered.
  • the object according to the invention is furthermore achieved by a preparation having the features of claim 11.
  • the preparation according to the invention comprises cells according to the invention, for example neural crest-derived cells, eg melanocyte progenitor cells, and / or keratinocyte precursor cells and / or mesenchymal progenitor cells.
  • the preparation is present as a suspension.
  • the preparation is suitable and / or intended to be applied directly to a body site, for example, as described in the o. G. PCT / EP 2008/007684 described effects.
  • the cells can thereby be obtained by plucking hair from the vital skin of the donor or by plucking or otherwise obtaining hair from a present skin biopsy of the donor.
  • the cells are obtained and intended for use without having previously been spent or spent in a culture to grow cells. This may apply up to the receipt of an order condition of the product.
  • the kit according to the invention optionally comprises a frame in some embodiments, and also, optionally, a template.
  • it comprises a hair follicle unit (A).
  • the hair follicle unit (A) comprises all or several articles of a group, e.g. at least one syringe, z. B. about 5 ml
  • At least one cannula eg about 40 mm, 18G at least one cell strainer
  • the kit according to the invention comprises in some embodiments a preparation unit (B).
  • the preparation unit (B) comprises all or several articles of a group which are e.g. at least one small syringe (e.g., two small syringes), e.g. At least one cannula (e.g., two cannulas), e.g. about 40 mm, 18G at least one sterile filter (for example two sterile filters)
  • At least one cannula e.g. B. about 80 mm, 14G
  • the kit according to the invention comprises, in some embodiments, an application unit (C).
  • the application unit (C) has all or several objects of a group, which: at least one pair of tweezers
  • At least one syringe e.g. about 2 ml
  • At least one cannula e.g. about 40mm, 18G
  • At least one spray bottle e.g. about 5 ml
  • kits comprises at least one spray head.
  • solutions, enzymes, etc. used in the application of the kit according to the invention are present in some embodiments according to the invention as ampoules with the following content:
  • Ampoule 1 PBS buffer e.g. about 5 ml (hair follicle collection solution)
  • Ampoule 2 PBS buffer e.g. about 5 ml (to release the dispase
  • Ampoule 4 PBS buffer with EDTA e.g. about 5 ml (to release the
  • Ampoule 5 trypsin e.g., about 2 mg (15,000 BAEE) plus
  • Hyaluronidase e.g., about 25 mg (7500 IU)
  • lyophilizate mixture e.g., about 25 mg (7500 IU)
  • Ampoule 7 PBS buffer e.g. about 2 ml (cell resuspension buffer).
  • the kit comprises at least the first two of the following four ampoules lw, 2w, 3w and 4w or these are used in the method according to the invention of some embodiments.
  • the ampoules contain:
  • Ampoule lw PBS buffer to collect or store the
  • Ampoule 2w enzyme mixture (collagenase + neutral protease) - lyophilisate
  • the enzyme lyophilisate present in the ampoule 2w is dissolved with part of the contents of the ampoule lw.
  • This enzyme solution is added to the hair follicles in a reaction vessel.
  • the contents of the reaction vessel are mixed by shaking or otherwise.
  • the reaction vessel is placed in the heating bath and incubated for 30 minutes at 35 ° C (temperature control), then removed from the heating bath and shaken gently for 4 minutes.
  • the liquid content of the reaction vessel (cell suspension) is transferred to another reaction vessel.
  • the hair remains behind.
  • the further reaction vessel is centrifuged, for example for 5 minutes at 700 g.
  • the supernatant fluid is removed in the further reaction vessel.
  • the remaining pellet in the other reaction vessel is carefully dissolved, for example, using the contents of the ampoule 3w.
  • the further reaction vessel is centrifuged again, for example for 5 minutes at 700 g. Subsequently, the supernatant is removed again. The pellet is dissolved, for example by means of the contents of ampoule 4w. The cell suspension is thus ready for use.
  • the method includes plucking hair follicles, not others; in certain According to embodiments of the invention, the method comprises using plucked hair follicles.
  • the vessel in which the hair follicles are located is closed, for example by means of a septum. In such embodiments, it may be provided to at least after the introduction of the hair follicles into the vessel in a
  • closed system or as a closed system.
  • count or contribute that liquids are transferred only via septa in the vessels used with the help of syringes, needles, sterile filters and the like, until the cell suspension is ready to use.
  • the method comprises enzymatic digestion in the vessel with the hair follicles, preferably under controlled conditions of temperature and / or duration.
  • the temperature range can be between 18 °, preferably from 20 ° C to the area in which the cells die off, about 45 ° C.
  • the method includes one or more of the following optional steps:
  • Centrifuging at 700 g, but also in a range e.g. from 150 g up to 2000 g, the time between centrifuging briefly and up to 20 or more minutes;
  • the washing steps can in principle be repeated several times; and receiving the cell pellet in the appropriate volume for the application of buffer, for example 0.5 to 5 ml, possibly with the addition of viscosity-increasing substances.
  • the method comprises transferring the cell suspension into a suitable vessel for the application, e.g. Spray bottle.
  • a suitable vessel for the application e.g. Spray bottle.
  • the application is also possible directly from the centrifugation vessel, e.g. Application by brush, also with the addition of viscosity-increasing substances.
  • venting cannulae are provided. In certain embodiments according to the invention, these are provided with sterile filters in order to ensure that the ambient air which penetrates by means of the venting cannulas is sterile.
  • the kit of the embodiments shown here contain different enzymatic digests.
  • a sequential digestion is carried out first with dispase and then with trypsin and hyaluronidase in a mixture.
  • Dispase digestion is preferably carried out without EDTA, the
  • Trypsin digestion preferably with EDTA.
  • enzymes can also be used, such as e.g. papain, elastase, pronase, AccutaseTM, collagenases, neutral proteases and other enzymes from the enzyme class EC 3 (hydrolases).
  • dispase may be replaced or supplemented with one or more other, preferably neutral, proteases
  • trypsin may be replaced or supplemented by one or more other proteases
  • hyaluronidase may be replaced with one or more other enzymes capable of cleaving hyaluronic acid.
  • these enzymes can be replaced by other enzymes, the extracellular matrix components, e.g. Collagen or elastin, such as collagenases or elastase at least partially replaced or supplemented.
  • the PBS buffer with EDTA can be used, for example, as a liquid in the form of a clear, colorless, odorless solution prepared according to Ph. Eur (European Pharmacopoeia in the current version), as described, for example, by Biochrom AG, Germany under the article number L21 13 is available in a 100 ml glass bottle.
  • the trypsin / hyaluronidase / EDTA solution may e.g. about 0.8% (EDTA, w / v).
  • the solution causes in particular a detachment of the cells from the hair shaft.
  • the detachment is preferably carried out at about 37 ° C. However, higher temperatures are possible in which the viability of the cells is still guaranteed, or lower, in which an activity of trypsin is still guaranteed.
  • the PBS buffer can be obtained, for example, from BioConcept, Switzerland in a 500 ml bottle in liquid form with a pH of 7.3 ⁇ 0.2 and an osmolarity of 285 ⁇ 10 as PBS without Ca / Mg with the article no. 3-05F29 (PBS1) or as PBS buffer with Ca / Mg under the article no. 8-05F00 (PBS2) as a sterile, colorless and clear liquid which can be stored at room temperature.
  • This PBS may be suitable for the preparation of M solution.
  • An advantage of the enzymatic detachment procedure is, in particular, that the potential use of enzymes has a detrimental effect on the treated cells and / or their alteration, i.a. Differentiation or genetic modification can be avoided.
  • the enzymatic detachment is therefore particularly gentle on the cells to be recovered.
  • An advantage achievable by means of some embodiments of the present invention is that the inventive success in producing the cell suspension without centrifugation can be achieved.
  • This advantageously makes it possible to carry out the invention without special apparatus design, even without connection to a voltage source.
  • This can be made possible, for example, by washing the hair follicles after the enzyme treatment described below.
  • Such predissolved cells remain at least for the most part on the hair and are only by their movement, for example by shaking, panning, by ultrasound or any other type of physical action or acceleration in the final volume of cell resuspension buffer detached. You can then directly into a suitable container, z. B. a Sprühfl, be transferred and applied.
  • An advantage obtainable by means of certain embodiments of the present invention lies in the closed nature of the system. In a very small space, the conditions of aseptic production are fulfilled or complied with. This results from the fact that the vessels in which the
  • Hair follicles or cells are located, are closed off from the environment. moreover The added buffers and enzymes can also be introduced germ-free.
  • Fig. 1 shows a template with symbols for guiding an implementation of the method according to the invention using a
  • the three-dimensional frame contained in some embodiments of the kit according to the invention is in such a way also contained in a kit in some embodiments
  • the frame can have arrow markings, as can be seen in FIG. 1 and the template 100 can also be seen. Based on the illustrated arrows, the user can easily orient in the following by following them in the direction of the arrow. A omission of individual steps of the procedure described below is therefore the user, because z. B. a past in the direction of arrow depression was not filled or used, or does not happen at all.
  • the ampoules contained in the package of the kit are inserted into the corresponding recesses of the frame. This lowers advantageous the risk of later confusing the individual ampoules.
  • a first ampoule is placed or plugged into a first opening 1, through which the lettering Amp 1 of the template can be seen.
  • a second ampoule is placed in a second opening 2 (Amp 2), a third ampoule is placed in a third opening 3 (Amp 3), a fourth ampoule is placed in a fourth opening 4 (Amp 4), a fifth ampoule is placed in a fifth aperture 5 (Amp 5) is placed, a sixth ampule is placed in a sixth aperture 6 (labeled Amp 6 on the template), and a seventh ampule is placed in a seventh aperture 7 (Amp 7).
  • the hair follicle unit (A) or package A is opened.
  • a cell strainer contained therein is placed in a first recess 11 of the rack.
  • the first recess 11 in the present example is the one which is closest to the ampoule 1 inserted into the first opening 1.
  • the recess 11 may - like each of the other wells - a shell or other container for receiving a liquid.
  • the contents of the first ampoule namely PBS buffer about 5 ml (referred to herein as hair follicle collection solution), are removed by means of a syringe also contained in the kit A of the kit and a cannula of the first ampoule and placed in the cell strainer which is now in the first one
  • a number of hair follicles preferably from about 50 to about 500, and most preferably about 250, hair follicles which may have been previously plucked, especially prior to all of the steps of using the kit described herein, are freed of dead hair material.
  • the dead hair material can be z. B. be cut with a scissors also present in the package A.
  • the hair follicles are given in their present state in the solution present in the cell strainer.
  • the preparation unit (B) or package B present in certain embodiments of the kit is opened.
  • the contents of the second ampoule namely about 5 ml of PBS buffer (for dispensing the dispase lyophilisate), which is or is in a second orifice 2, are injected by means of a syringe (for example about 5 ml) and a syringe (for example about 40 mm) long) cannula added to lyophilisate, which is located in a third ampoule, which is in a third opening 3 and Dispase contains about 5 mg (5 U) of lyophilisate.
  • a syringe for example about 5 ml
  • a syringe for example about 40 mm
  • the contents of the third ampoule are subsequently re-wound up by means of the syringe.
  • the cannula is removed from the filled syringe. Instead of the cannula, a sterile filter is placed on the syringe.
  • the syringe contents are passed out of the syringe through the sterile filter into the second depression 12, which in the example of FIG. 1 now lies closest to the third ampule.
  • the cell strainer which has the hair follicles, is removed from the first recess 11 and placed in a second recess 12 such or rather such that the cell strainer passive, d. h without the support of the User, z. B. due to its own weight, can dip into the second recess 12.
  • the tweezers attached to the kit in some embodiments may be used.
  • the cell sieve is allowed to stand in the second recess 12 for a first waiting time of preferably about 10 to 20 minutes, in particular about 15 minutes.
  • the contents of the fourth vial containing PBS buffer with EDTA, about 5 ml (to dissolve the trypsin / hyaluronidase lyophilizate mixture) are mixed with a second (eg 5 ml) syringe and a second
  • cannula both included in the kit, can be added to lyophilisate in the fifth vial containing trypsin, about 2 mg (15,000 BAEE), and hyaluronidase, about 25 mg (7500 IU), lyophilizate mixture ,
  • trypsin about 2 mg (15,000 BAEE
  • hyaluronidase about 25 mg (7500 IU)
  • lyophilizate mixture a fifth vial containing trypsin, about 2 mg (15,000 BAEE), and hyaluronidase, about 25 mg (7500 IU), lyophilizate mixture .
  • the second cannula is removed from the filled second syringe. Instead, a sterile filter is placed on the second syringe.
  • Syringe contents are passed through the sterile filter into a third depression 13 which is closest to the fifth ampule.
  • the cell sieve with the hair follicles is lifted out of the second depression 12 and passively inserted into or onto the third depression 13 in such a way that the cell sieve can dip into the third depression 13.
  • the cell strainer is left a second waiting time, preferably about 30 minutes.
  • wash buffer from the sixth ampoule containing PBS buffer with Mg ions, about 30 ml (referred to herein as wash buffer) is mixed with one (e.g., about 30 ml syringe and a (for example, about 80 mm long) cannula, using a (for example about 25 mm wide) ventilation cannula, about 30 ml solution taken and each about 5 ml of it in the six wells provided for this purpose 14, 15, 16, 17, 18 and 19 distributed.
  • one e.g., about 30 ml syringe and a (for example, about 80 mm long) cannula, using a (for example about 25 mm wide) ventilation cannula, about 30 ml solution taken and each about 5 ml of it in the six wells provided for this purpose 14, 15, 16, 17, 18 and 19 distributed.
  • the cell strainer with the hair follicles is lifted out of the third recess 13 to be successively gently immersed in the now filled six wells 14, 15, 16, 17, 18 and 19 passively and lifted out of this again.
  • the application unit (C) or package C is opened.
  • the seventh vial, containing PBS buffer, about 2 ml (referred to herein as cell resuspension buffer), is opened by lifting its rubber stopper or lid.
  • the cell strainer is placed in the last well, the well 20.
  • the hair follicles are carefully transferred from the cell strainer into the seventh ampule using tweezers.
  • the seventh ampoule is closed again with the rubber stopper.
  • the seventh vial is then gently shaken for preferably about two to about six minutes, more preferably about four minutes.
  • the container present in the kit in some embodiments for example a vial which, for example, holds about 5 ml volume and may, for example, have a screw cap (eg a spray vial), is placed in the eighth opening 8 provided in the frame for this purpose.
  • the eighth opening 8 is above a corresponding marking, eg "spray", the template.
  • the contents of the seventh vial 7 are transferred into the spray vial using a syringe (for example, about 2 ml) and a cannula (for example, about 40 mm long). This process is done gently and slowly so as not to damage the cells in the solution.
  • the hair remains in the seventh ampule 7.
  • a spray head is then placed on the spray bottle or screwed, or this is closed in other ways.
  • the cell suspension is ready to be sprayed onto a body site to be treated.
  • the cell resuspension buffer additionally contains hyaluronic acid (for example about 0.3%), which advantageously contributes to increasing the viscosity.
  • hyaluronic acid instead of hyaluronic acid, it is of course also possible to use any other viscosity-increasing substance which has no adverse effect on the success to be achieved according to the invention.
  • the kit may alternatively or additionally have a brush.
  • Brushing or spraying but also be provided for dabbing or the like and / or applied accordingly.
  • centrifugation is not provided in some embodiments of the method according to the invention.
  • the method can thus proceed in such embodiments without centrifuging. This can keep the apparatus required, which is necessary for carrying out the method, advantageously low.
  • the further embodiment according to the invention can, apart from the plucking of the hair follicles, to minimize the germ load in one
  • Ampoules can remain closed. The addition or removal of
  • Liquids are made by septa, which are present in the lids of the reaction vessels and / or ampoules.
  • the septa are to be disinfected before piercing as possible.
  • Every new step involves working with fresh cannulas and syringes. It is always to work with gloves.
  • a temperature of 35 ° C is set in a heating bath.
  • the rack is - as far as provided - unpacked for containers and instruments to be used.
  • the following four ampoules lw, 2w, 3w and 4w are used. They contain:
  • the reaction vessel 1 is opened and placed in the rack. About 3 ml of the contents of ampoule lw is added to the reaction vessel 1.
  • a number of hair follicles preferably from about 50 to about 500 and most preferably about 250 hair follicles, are picked. Dead hair material will be cut off. The hair follicles are placed in the reaction vessel 1. Care is taken to ensure that all hair follicles in the
  • reaction vessel 1 is placed in the heating bath and incubated for 30 minutes at 35 ° C (temperature control), then removed from the heating bath and shaken gently for 4 minutes.
  • Reaction vessel 2 is centrifuged for 5 minutes at 700 g.
  • the pellet is carefully dissolved by the contents of the ampoule 3 w.
  • the reaction vessel 2 is again centrifuged for 5 minutes at 700 g.
  • Ampoule 1 ampoule lw:
  • Ampoule 3 Ampoule 2w:
  • Dispase-lyophilisate enzyme mixture (collagenase + neutral
  • Ampoule 6 Ampoule 3w:
  • Ampoule 7 Ampoule 4w:
  • the enzyme digests have to be carried out separately, the enzymes in the further embodiment can advantageously be used together since trypsin requires others
  • Buffers and possible additives This applies in principle to the procedure described for FIG. 1 and for the further embodiment. According to the invention, in both cases, by a specific combination or selection of the used Enzymes some or the enzymatic digestion are carried out together or at the same time.

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  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biomedical Technology (AREA)
  • Genetics & Genomics (AREA)
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  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
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  • Chemical & Material Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Dermatology (AREA)
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  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Cell Biology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

L'invention concerne un procédé de production d'une préparation, en particulier d'une suspension, qui contient des cellules du corps humain. L'invention concerne en outre une préparation et un kit. Ledit procédé comprend : une dissolution enzymatique de cellules du follicule pileux humain ; un lavage des cellules dissoutes et une mise en suspension des cellules dans une solution tampon pour obtenir une préparation. Un mode de réalisation est mis en œuvre de préférence sans étape de centrifugation avec des instruments faciles à se procurer, un deuxième mode de réalisation est mis en ouvre de préférence sous la forme d'un système fermé pour empêcher l'introduction de germes pendant la mise en œuvre.
PCT/EP2012/000657 2011-02-15 2012-02-15 Procédé de production d'une préparation en utilisant des cellules du follicule pileux, préparation et kit Ceased WO2012110232A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP12705789.1A EP2768947A1 (fr) 2011-02-15 2012-02-15 Procédé de production d'une préparation en utilisant des cellules du follicule pileux, préparation et kit

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
DE102011011309.6 2011-02-15
DE102011011309 2011-02-15
US201161468583P 2011-03-29 2011-03-29
DE102011011623A DE102011011623A1 (de) 2011-03-29 2011-03-29 Tischfußballspiel
DE102011011623.6 2011-03-29
US61/468,583 2011-03-29

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WO2012110232A1 true WO2012110232A1 (fr) 2012-08-23

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007100870A2 (fr) * 2006-02-28 2007-09-07 The Trustees Of Columbia University In The City Of New York Procédés d'agrégation compacte de cellules dermiques
WO2008004819A1 (fr) * 2006-07-05 2008-01-10 Seoul National University Industry Foundation Procédé pour isoler une cellule souche de follicule pileux et composition pour la reproduction des cheveux
EP1950284A1 (fr) * 2005-09-30 2008-07-30 Phoenixbio Co., Ltd. Procede de culture de cellules de la cuticule d'un follicule pileux

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1950284A1 (fr) * 2005-09-30 2008-07-30 Phoenixbio Co., Ltd. Procede de culture de cellules de la cuticule d'un follicule pileux
WO2007100870A2 (fr) * 2006-02-28 2007-09-07 The Trustees Of Columbia University In The City Of New York Procédés d'agrégation compacte de cellules dermiques
WO2008004819A1 (fr) * 2006-07-05 2008-01-10 Seoul National University Industry Foundation Procédé pour isoler une cellule souche de follicule pileux et composition pour la reproduction des cheveux

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
SEO YOUNG-KWON ET AL: "Development of isolation and cultivation method for outer root sheath cells from human hair follicle and construction of bioartificial skin.", BIOTECHNOLOGY AND BIOPROCESS ENGINEERING, vol. 8, no. 2, March 2003 (2003-03-01), pages 151 - 157, XP002673905, ISSN: 1226-8372 *

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