JP2003235990A - Method for discharging minute organismic material - Google Patents

Method for discharging minute organismic material

Info

Publication number
JP2003235990A
JP2003235990A JP2002035945A JP2002035945A JP2003235990A JP 2003235990 A JP2003235990 A JP 2003235990A JP 2002035945 A JP2002035945 A JP 2002035945A JP 2002035945 A JP2002035945 A JP 2002035945A JP 2003235990 A JP2003235990 A JP 2003235990A
Authority
JP
Japan
Prior art keywords
papilla
solution
cells
tissue
fine
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.)
Pending
Application number
JP2002035945A
Other languages
Japanese (ja)
Inventor
Kimie Toyoshima
公栄 豊島
Katsutoshi Yoshizato
勝利 吉里
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.)
Institute of Immunology Co Ltd
Japan Science and Technology Agency
Hiroshima Industrial Promotion Organization
Original Assignee
Institute of Immunology Co Ltd
Hiroshima Industrial Promotion Organization
Japan Science and Technology Corp
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
Application filed by Institute of Immunology Co Ltd, Hiroshima Industrial Promotion Organization, Japan Science and Technology Corp filed Critical Institute of Immunology Co Ltd
Priority to JP2002035945A priority Critical patent/JP2003235990A/en
Publication of JP2003235990A publication Critical patent/JP2003235990A/en
Pending legal-status Critical Current

Links

Landscapes

  • Media Introduction/Drainage Providing Device (AREA)
  • Materials For Medical Uses (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for surely and stably discharging a specified amount of a minute organismic material from a discharge device such as an injector. <P>SOLUTION: After a polymer polysaccharide solution is brought into contact with the inside wall surface of the liquid discharge device, liquid including the minute organismic material is sucked and discharged into and from the discharge device. <P>COPYRIGHT: (C)2003,JPO

Description

【発明の詳細な説明】Detailed Description of the Invention

【0001】[0001]

【発明の属する技術分野】この出願の発明は、細胞、組
織片または人工組織片等の微細な生体材料を含む液体
を、液体排出装置の内壁に付着残留させることなく、一
定量を確実かつ安定的に排出させることを目的とする技
術に関するものである。
BACKGROUND OF THE INVENTION 1. Field of the Invention The invention of this application is to ensure a certain amount of a liquid containing fine biomaterial such as cells, tissue pieces or artificial tissue pieces, without sticking and remaining on the inner wall of the liquid discharge device. The present invention relates to a technology for the purpose of discharging it.

【0002】[0002]

【従来の技術とその課題】毛髪を作り出す毛包組織は、
胎児期においてパピラ(毛乳頭)と呼ばれる特殊な間充
織と表皮との相互作用によって形成される。また、パピ
ラは毛髪の伸長と休止の周期である毛周期の進行にも深
く関与している。種々の原因によってこの毛周期が変調
を来すことで、男性型脱毛症が発症する。男性型脱毛症
の後期ではパピラそのものが死滅減少するため発毛治療
薬等の効果は低く、さらに、放射線障害による脱毛や円
形脱毛症では有効な毛包の再生による発毛治療も、男性
型脱毛症では期待できない。
2. Description of the Related Art Hair follicle tissue that produces hair is
It is formed by the interaction between the epidermis and a special mesenchyme called papilla in the fetal period. Papilla is also deeply involved in the progression of the hair cycle, which is the cycle of hair elongation and rest. Modulation of this hair cycle due to various causes causes male pattern baldness. In the latter stage of androgenetic alopecia, the effects of hair growth remedies and the like are low because the papilla itself is diminished and death.In addition, hair loss due to radiation damage or hair loss by alopecia areata is effective for male hair loss. I can't expect it from the disease.

【0003】したがって、脱毛症の治療では、毛包誘導
能力を持つパピラの移植治療が最も有効なものとして期
待されている。パピラは後頭部あるいは側頭部の脱毛症
に罹患していない部位から摘出することができる。
Therefore, in the treatment of alopecia, transplantation treatment of papilla having a hair follicle-inducing ability is expected to be the most effective. The papilla can be removed from a region of the occipital region or temporal region not affected by alopecia.

【0004】従来、パピラを移植する場合には、まず鋭
利なピンセットやメスによって患者皮膚を切開し、摘出
したパピラを手作業で真皮・表皮間隙へ挿入していた。
この移植によって、移植創に毛包構造や毛髪が誘導され
る(Development, 122, 3085-3094, 1996)。
Conventionally, when a papilla is transplanted, first, the patient's skin is incised with sharp tweezers or a scalpel, and the extracted papilla is manually inserted into the dermis / epidermis gap.
This transplantation induces hair follicle structure and hair in the transplant wound (Development, 122, 3085-3094, 1996).

【0005】しかしながら、こういった手作業で移植を
行うには特殊な技能と多大な労力を必要とし、さらに移
植部位の皮膚へ大きなダメージを与えることになる。こ
のため、脱毛症患者の治療に必要な数千〜数万本の発毛
を促すために、毛髪と同数のパピラを移植することは実
質上不可能であった。そこで、注射器等の注入装置を用
いてパピラを頭皮に注入することも試みられているが、
その場合には、注入筒内壁へパピラが付着してしまい、
排出効率が著しく低下するといった問題点が存在した。
However, performing such manual transplantation requires special skill and a great deal of labor, and further causes great damage to the skin at the transplant site. Therefore, it is virtually impossible to transplant as many papilla as the number of hairs in order to promote hair growth of thousands to tens of thousands required for treatment of alopecia patients. Therefore, it has been attempted to inject papilla into the scalp using an injection device such as a syringe.
In that case, the papilla adheres to the inner wall of the injection cylinder,
There was a problem that the emission efficiency was significantly reduced.

【0006】また、このような組織付着の問題は、パピ
ラ以外の微細生体組織や細胞溶液の一定量を排出して分
配しようとする場合も同様であった。
Further, the problem of such tissue adhesion is the same as in the case of discharging and dispensing a certain amount of a fine living tissue other than papilla or a cell solution.

【0007】この出願の発明は、以上の通りの従来技術
の問題点に鑑みてなされたものであって、細胞は組織片
等の微細生体材料を、注射器等の排出装置から確実かつ
安定的に排出するための方法を提供することを課題とし
ている。
The invention of this application was made in view of the problems of the prior art as described above, and cells reliably and stably remove a fine biomaterial such as a tissue piece from an ejector such as a syringe. The challenge is to provide a method for emissions.

【0008】[0008]

【課題を解決するための手段】この出願は、前記の課題
を解決するための発明として、以下の工程: (1) 液体排出装置の内壁面に高分子多糖溶液を接触さ
せる工程; (2) 微細生体材料を含む液体を排出装置内へ吸入する
工程;および (3) 排出装置より液体を排出する工程、を有すること
を特徴とする微細生体材料の排出方法を提供する。
[Means for Solving the Problems] This application is an invention for solving the above problems and comprises the following steps: (1) a step of bringing a high molecular polysaccharide solution into contact with the inner wall surface of a liquid discharge device; (2) A method for discharging a fine biomaterial, comprising: a step of sucking a liquid containing the fine biomaterial into the discharge device; and (3) a step of discharging the liquid from the discharge device.

【0009】この方法においては、微細生体材料が、細
胞、組織片または人工組織片であること、さらに具体的
には、細胞がパピラ細胞を含む細胞混合物、組織片がパ
ピラ、人工組織片が培養パピラ細胞から形成した人工パ
ピラであることを好ましい態様としている。
In this method, the fine biomaterial is cells, tissue pieces or artificial tissue pieces, and more specifically, cells are a cell mixture containing papilla cells, tissue pieces are papilla, and artificial tissue pieces are cultured. A preferred embodiment is an artificial papilla formed from papilla cells.

【0010】[0010]

【発明の実施の形態】この出願の発明において、排出す
る微細生体材料は、生体を構成する各種の細胞、細胞や
タンパク質等によって構成される生体組織片、または培
養細胞から人工的に構築された人工組織片である。細胞
は特段の制限はなく、また細胞の排出用途も移植に限定
されることはない。例えば培養細胞の所定量を別の培地
に分配するためなどに使用することができる。組織片も
由来臓器に制限はなく、例えば歯乳頭のような移植組織
片も対象とすることができる。人工組織片も、例えば、
特開平2001-340076号公報や特開表09-510108号公報等に
開示されているような、培養細胞から構築した移植用人
工組織の微細片を使用することができる。
BEST MODE FOR CARRYING OUT THE INVENTION In the invention of this application, the fine biomaterial to be discharged is artificially constructed from various cells constituting a living body, biological tissue pieces composed of cells and proteins, or cultured cells. It is an artificial tissue piece. The cells are not particularly limited, and the cell excretion use is not limited to transplantation. For example, it can be used for distributing a predetermined amount of cultured cells to another medium. The tissue piece is not limited to the organ of origin, and a transplanted tissue piece such as a dental papilla can also be a target. Artificial tissue pieces, for example,
Fine pieces of artificial tissue for transplantation constructed from cultured cells as disclosed in JP-A-2001-340076 and JP-A-09-510108 can be used.

【0011】さらに、この発明の方法では、細胞として
パピラ細胞を使用することを好ましい態様の一つとして
いる。パピラ細胞は、例えば、特開平7-274950号公報に
記載の方法で頭皮から単離したものを使用することがで
きる。そして、このパピラ細胞を移植用に使用する場合
には、パピラ細胞と表皮細胞、線維芽細胞との混合細胞
(特願2002-035815号:出願日2002年2月13日)を含む液
体(細胞浮遊液)を排出の対象とすることもできる。
Furthermore, in the method of the present invention, the use of papilla cells as cells is one of the preferred embodiments. As the papilla cells, those isolated from the scalp by the method described in JP-A-7-274950 can be used, for example. When the papilla cells are used for transplantation, a liquid (cells containing mixed cells of papilla cells, epidermal cells, and fibroblasts (Japanese Patent Application No. 2002-035815: filing date February 13, 2002)) Suspension liquid) can also be targeted for discharge.

【0012】この発明の方法はまた、組織片としてパピ
ラを使用することを別の好ましい態様としている。パピ
ラは、公知の方法によってヒト頭皮より無菌状態で実体
顕微鏡下にて単離し、このパピラを血清や培地等の適当
な担体溶液に懸濁したもの(パピラ浮遊液)を使用する
ことができる。
[0012] The method of the present invention also makes use of papilla as the tissue piece another preferred embodiment. The papilla can be isolated from a human scalp by a known method in a sterile state under a stereomicroscope, and the papilla suspended in an appropriate carrier solution such as serum or medium (papilla suspension) can be used.

【0013】さらにまたこの発明の方法では、人工組織
片として、培養パピラ細胞から構築した人工パピラを使
用することをさらに別の態様としている。この人工パピ
ラは、特開平7-274950号公報に記載の方法により継代培
養したパピラ細胞を、例えばスフェロイドプレート(住
友ベークライト製)等の培養基材に任意の数播種し、50
% CM5等を含むDMEM10培地等で24時間程度培養すること
によって構築することができる。
Furthermore, in the method of the present invention, as another embodiment, an artificial papilla constructed from cultured papilla cells is used as the artificial tissue piece. This artificial papilla is prepared by subculturing papilla cells subcultured by the method described in Japanese Patent Laid-Open No. 7-274950 into an arbitrary number, for example, on a culture substrate such as a spheroid plate (Sumitomo Bakelite Co., Ltd.), 50
It can be constructed by culturing in DMEM10 medium containing% CM5 etc. for about 24 hours.

【0014】この発明の方法は、前記のとおりの微細生
体材料を液性担体(例えば、血清や培養液等)に混合
し、注射器等の液体排出装置に吸入した後、一定量を排
出する。その際に、この発明の方法は、事前に液体排出
装置の内壁(例えば、注射筒の内壁)に高分子多糖のゲ
ル状溶液を接触させる。高分子多糖は、カルボキシル基
あるいは硫酸基にて陰性荷重を与えられた、例えば粘度
の高いカルボキシメチルセルロース(CMC)や、ヒアル
ロン酸、ヘパラン硫酸、デルマタン硫酸、ケタラン硫
酸、コンドロイチン硫酸等、およびこれらの塩を使用す
ることができる。これらの高分子多糖は、適当な液体、
好ましくは微細生体材料との混合液として使用する液体
と同一の液体に、0.1〜10%(W/V)程度の濃度で混合
し、溶液化する。そして、この溶液を、液体排出装置に
吸入し、装置内壁面を高分子多糖溶液でコーティングし
た後、微細生体材料液を液体排出装置に充填する。すな
わち、排出装置の内壁は、陰性電荷を持ち粘性の高い高
分子多糖類でコーティングされているため、粘性の低い
血清や培地等からなる微細生体材料液とは容易に混じら
ず、微細生体材料が排出装置内壁と直接接触することは
ない。これによって、微細生体材料は排出装置の内壁に
付着残存することがないため、微細生体材料を予め決定
した濃度のまま正確に排出することが可能となる。また
連続的に一定量を排出することができるため、装置の自
動化も可能となる。なお、事前に吸入した高分子多糖溶
液は、微細生体材料液を吸入する前に排出させてもよ
く、あるいは高分子多糖溶液を排出せずに、引き続き微
細生体材料液を吸入してもよい。特に後者の場合は、そ
れぞれの液体が粘性の違いにより混合することがないた
め、たとえ排出装置内に高分子多糖溶液が残存した場合
であっても、微細生体材料溶液のみが排出される。
In the method of the present invention, the fine biomaterial as described above is mixed with a liquid carrier (for example, serum or culture solution), which is sucked into a liquid discharging device such as a syringe and then discharged in a fixed amount. At that time, in the method of the present invention, the gelled solution of the high molecular polysaccharide is brought into contact with the inner wall of the liquid discharge device (for example, the inner wall of the syringe) in advance. High molecular weight polysaccharides are negatively loaded with a carboxyl group or a sulfate group, for example, highly viscous carboxymethyl cellulose (CMC), hyaluronic acid, heparan sulfate, dermatan sulfate, ketalane sulfate, chondroitin sulfate, and salts thereof. Can be used. These high molecular polysaccharides are suitable liquids,
Preferably, the same liquid as the liquid used as the mixed liquid with the fine biomaterial is mixed at a concentration of about 0.1 to 10% (W / V) to form a solution. Then, this solution is sucked into the liquid discharging device, the inner wall surface of the device is coated with the polymer polysaccharide solution, and then the liquid discharging device is filled with the fine biomaterial liquid. That is, since the inner wall of the discharging device is coated with a high-molecular polysaccharide having a negative charge and high viscosity, it does not easily mix with a fine biomaterial liquid such as serum or medium having low viscosity, and the fine biomaterial is There is no direct contact with the inner wall of the ejector. As a result, since the fine biomaterial does not adhere to and remain on the inner wall of the discharging device, it is possible to accurately discharge the fine biomaterial in a predetermined concentration. Further, since a fixed amount can be continuously discharged, the device can be automated. The polymer polysaccharide solution previously inhaled may be discharged before inhaling the fine biomaterial solution, or the fine biomaterial solution may be continuously inhaled without discharging the polymer polysaccharide solution. Especially in the latter case, since the respective liquids do not mix due to the difference in viscosity, even if the high molecular polysaccharide solution remains in the discharging device, only the fine biomaterial solution is discharged.

【0015】以下、実施例を示してこの出願の発明につ
いてさらに詳細かつ具体的に説明するが、この出願の発
明は以下の例に限定されるものではない。
Hereinafter, the invention of this application will be described in more detail and specifically with reference to Examples, but the invention of this application is not limited to the following examples.

【0016】[0016]

【実施例】1. 方法 (a) 頭皮組織からのパピラの単離 被験者頭部より切除した頭皮組織はイソジン液、70%ア
ルコールによって消毒した。保存する場合は、10%被験
者血清あるいは牛胎児血清を含むダルベッコ改変イーグ
ル培地(DMEM10)を用いた。パピラは注意深く真皮性毛
根鞘および毛母から無菌的に分離し、被験者の血清中で
保存した。 (b) 高分子多糖溶液の調製 カルボキシメチルセルロースナトリウム(CMC-Na)を3.
5%となるように無血清DMEM培地へ懸濁し、一晩20rpmで
攪拌しつつ水和させた。水和したCMC-Na溶液は121℃、1
5分間の条件で高圧蒸気滅菌を施し、使用まで冷蔵し
た。また同様に1%のヒアルロン酸ナトリウム溶液(生化
学工業株式会社製)を無菌環境下で使用した。 (c) 注射筒の準備 パピラ注入用注射筒は、内容量100μlのマイクロシリン
ジ(HAMILTON社製)を用い、高圧蒸気滅菌後に使用し
た。マイクロシリンジ内にCMC-Na溶液を40μl吸入し
た。次いで、20mM HEPESを含む無血清DMEM培地(pH 7.
5)を20μl吸入した。マイクロシリンジの注射針接合部
を滅菌ガーゼでよく拭き取り、27G注射針を取り付け、
無血清DMEM培地中に注射針先端を浸し、ピストンを注意
深く押し出して、マイクロシリンジおよび注射針内の残
留空気を無血清DMEM培地およびCMC-Na溶液と共に完全に
押し出し、シリンジ内に20μlのCMC-Na溶液を残存させ
た。次に、被験者自身より採取した血清に浮遊させたパ
ピラ3個(全量80μl以内)を注射筒に吸入した。ヒア
ルロン酸ナトリウム溶液をした場合も同様とした。 (d) パピラの膨留疹形成術を伴う皮内注射による移植 注射筒内にパピラを装填した後、速やかに被験者皮膚へ
移植を行った。被験者皮膚は予め下腕部内側に表面麻酔
を施し、痛覚が消失したことを確認した後に10箇所の下
腕部内側皮膚へ移植術を施した。27G注射針は移植目標
部位より5mm離れた位置より皮膚に対してほぼ平行に表
皮直下の真皮層内へ刺入し膨留疹を形成させた。このと
き、注射筒内に少なくとも20μlの液体を残すことで、
高分子多糖溶液の皮膚内への漏出を防止した。また同様
に、ヌードマウス背部にDiI蛍光色素標識したラット頬
髭パピラを移植し、CMC-Na溶液の注入効率を無血清DMEM
培地あるいは血清のみと比較評価した。さらにヌードマ
ウスへの移植10日目に、移植部位の皮膚を摘出し、通法
に従ってパラフィン切片として蛍光色素で標識された移
植パピラを追跡した。 (e) パピラ移植による発毛観察 移植前に顕微鏡観察し、移植部位に体毛の有無を確認し
た。移植後1〜2週間ごとに顕微鏡観察し移植皮膚の変
化を観察した。 2. 結果 陰性電荷を持つ高分子多糖類溶液と、血清に浮遊したパ
ピラの注射筒および注射針内での挙動を図1Aに図示す
る。図1A-1から図1A-4までがパピラ装填までの挙動であ
り、図1A-5から図1A-7がパピラ排出時の様子を示す。CM
C-Na溶液(ゲル)またはヒアルロン酸ナトリウム溶液
(ゲル)を充填したマイクロシリンジに血清中に浮遊さ
せたパピラを吸入すると、図1A-1〜4に示した様に、高
分子多糖溶液内部に形成される管上の経路をたどって、
パピラはシリンジのピストン側へ移行し、血清層に留ま
る(図1B、C)。
[Examples] 1. Method (a) Isolation of papilla from scalp tissue The scalp tissue excised from the subject's head was disinfected with isodine solution, 70% alcohol. For storage, Dulbecco's modified Eagle medium (DMEM10) containing 10% subject serum or fetal calf serum was used. Papilla was carefully aseptically separated from the dermal root sheath and hair matrix and stored in the serum of the subjects. (b) Preparation of high-molecular polysaccharide solution Add sodium carboxymethyl cellulose (CMC-Na) to 3.
The suspension was suspended in serum-free DMEM medium at 5% and hydrated overnight with stirring at 20 rpm. Hydrated CMC-Na solution at 121 ° C, 1
High-pressure steam sterilization was performed under the condition of 5 minutes and refrigerated until use. Similarly, a 1% sodium hyaluronate solution (manufactured by Seikagaku Corporation) was used under a sterile environment. (c) Preparation of Injection Cylinder A syringe for injection of papilla was a microsyringe (manufactured by HAMILTON) having an internal volume of 100 μl, and was used after autoclaving. 40 μl of CMC-Na solution was inhaled into the microsyringe. Then, serum-free DMEM medium containing 20 mM HEPES (pH 7.
20 μl of 5) was inhaled. Wipe the injection needle joint of the micro syringe well with sterile gauze, attach the 27G injection needle,
Immerse the needle tip in serum-free DMEM medium and carefully push out the piston to completely push out residual air in the microsyringe and needle together with serum-free DMEM medium and CMC-Na solution, and 20 μl CMC-Na in the syringe. The solution remained. Next, 3 papilla suspended in the serum collected from the subject himself (total amount within 80 μl) was inhaled into the syringe. The same applies when a sodium hyaluronate solution is used. (d) Transplantation of papilla by intracutaneous injection accompanied by blister plasty After the papilla was loaded into the syringe barrel, the skin of the subject was immediately transplanted. The subject's skin was previously subjected to surface anesthesia on the inside of the lower arm, and after confirming that the pain sensation had disappeared, transplantation was performed on the skin of the lower arm at 10 locations. The 27G injection needle was inserted into the dermis layer just below the epidermis at a distance of 5 mm from the target site for implantation and formed a rash. At this time, by leaving at least 20 μl of liquid in the syringe,
The leakage of the high molecular polysaccharide solution into the skin was prevented. Similarly, a rat whiskers papilla labeled with DiI fluorescent dye was transplanted to the back of a nude mouse, and the injection efficiency of CMC-Na solution was adjusted to serum-free DMEM.
Comparative evaluation was performed with only the medium or serum. Furthermore, on the 10th day from the transplantation into nude mice, the skin at the site of transplantation was excised, and the paraffin section-transplanted papilla labeled with a fluorescent dye was traced according to a conventional method. (e) Hair growth observation by papilla transplantation Before the transplantation, microscopic observation was performed to confirm the presence or absence of hair at the transplant site. Every 1-2 weeks after the transplantation, microscopic observation was performed to observe changes in the transplanted skin. 2. Results The behavior of high-molecular-weight polysaccharide solution with negative charge and papilla suspended in serum in the syringe barrel and needle is shown in Fig. 1A. 1A-1 to 1A-4 show the behavior up to loading the papilla, and FIGS. 1A-5 to 1A-7 show the behavior at the time of discharging the papilla. cm
When a papilla suspended in serum was inhaled into a microsyringe filled with a C-Na solution (gel) or a sodium hyaluronate solution (gel), as shown in Figs. Follow the path on the tube that is formed,
The papilla migrates to the piston side of the syringe and stays in the serum layer (Fig. 1B, C).

【0017】CMC-Naゲルを用いた場合の注入効率と用い
ない場合の注入効率をヌードマスウ背部皮膚にラット頬
髭を移植して比較した。それぞれの注入率は、注射針か
ら排出されたパピラの数によって求めた。その結果、表
1に示したように、血清を含まないDMEM培地または血清
のみでは、シリンジ内または注射針内にパピラが付着残
留するため、CMC-Na溶液を用いた注入法に比べ、注入効
率が著しく低かった。
The injection efficiency with and without CMC-Na gel was compared by implanting rat whiskers on the back skin of nude mice. The injection rate of each was determined by the number of papillas ejected from the injection needle. As a result, as shown in Table 1, in the serum-free DMEM medium or serum alone, papilla adhered and remained in the syringe or the injection needle, so that the injection efficiency was higher than that of the injection method using the CMC-Na solution. Was remarkably low.

【0018】[0018]

【表1】 また、実際にヒト皮膚への移植においてもCMC-Na溶液を
注射筒内に事前に吸入した場合には、表2に示したよう
に、83.3〜100%という高い注入率を得た。移植後の皮膚
は図3Aで示すような膨留疹が形成されるが、3週間ほど
で消失した。また、同様にヒアルロン酸ナトリウム(Hy
al. Na)を用いた場合でも、表2のとおりに73.3%およ
び76.7%という高い注入率が得られた。さらに、表2に
示した症例番号1、4、5番において新生毛が観察され
た。発毛率はそれぞれパピラの移植数に対して8.3%、1
0.0%、3.3%であった。また移植後に発生した新生毛は図
3B〜Dのように一カ所から2〜4本発生した。またその色
は白色と黒色両方とも観察された。
[Table 1] Further, even when the CMC-Na solution was actually inhaled into the syringe even when it was actually transplanted to human skin, as shown in Table 2, a high injection rate of 83.3 to 100% was obtained. A swollen rash was formed on the skin after transplantation as shown in FIG. 3A, but it disappeared in about 3 weeks. Similarly, sodium hyaluronate (Hy
al. Na), high injection rates of 73.3% and 76.7% were obtained as shown in Table 2. Furthermore, new hair was observed in case numbers 1, 4, and 5 shown in Table 2. Hair growth rates were 8.3% and 1 for papilla transplants, respectively.
It was 0.0% and 3.3%. In addition, the new hair generated after transplantation is
As in 3B to D, 2 to 4 occurred from one place. The color was observed both white and black.

【0019】[0019]

【表2】 ヌードマウス皮膚における移植パピラを組織学的に追跡
したところ、CMC-Na溶液を用いた膨留疹形成術では、ヌ
ードマウス皮膚の真皮層にパピラが注入されていること
が確認された(図2C〜H)。
[Table 2] Histological follow-up of the transplanted papilla in the skin of nude mice confirmed that papula was injected into the dermal layer of nude mouse skin in the ulcer formation procedure using a CMC-Na solution (Fig. 2C). ~ H).

【0020】[0020]

【発明の効果】以上詳しく説明した通り、この出願の発
明によって、一定量の微細生体材料を確実かつ安定的に
排出することが可能となる。これによって、例えばパピ
ラ等の移植による脱毛症治療が可能となる。
As described in detail above, according to the invention of this application, it becomes possible to reliably and stably discharge a certain amount of fine biomaterial. This enables alopecia treatment by transplantation of papilla, for example.

【図面の簡単な説明】[Brief description of drawings]

【図1】高分子多糖溶液としてCMC-Naゲルを用い、移植
組織をマイクロシリンジ内に吸入、排出した状態を示
す。A1-7はマイクロシリンジ内のCMC-Naゲルと移植組織
との挙動を連続的に示した模式図、Bは実際にCMC-Naお
よびパピラ/血清をマイクロシリンジに充填した写真で
ある。CはBの拡大写真であり、白い毛乳頭がシリンジ内
の血清層に存在している(矢印)。
FIG. 1 shows a state in which CMC-Na gel is used as a polymer polysaccharide solution, and the transplanted tissue is inhaled and discharged into a microsyringe. A1-7 is a schematic view showing continuously the behavior of CMC-Na gel in the microsyringe and the transplanted tissue, and B is a photograph in which CMC-Na and papilla / serum were actually filled in the microsyringe. C is a magnified photograph of B, with white papillae present in the serum layer in the syringe (arrow).

【図2】この発明の方法によってパピラをマイクロシリ
ンジから排出し、ヌードマウスの皮内に注射した写真像
である。Aはあらかじめ蛍光色素DiIによって標識したパ
ピラ、Bは移植箇所を示したヌードマウス、Cは皮内移植
から10日目の移植部位組織観察像(HE染色)、Dは他の
移植部位組織像(HE染色)である。EとFはそれぞれCとD
に示した組織の連続切片の核染色像、GとHは同じくDiI
蛍光像である。スケールバーは100μmを示す。
FIG. 2 is a photographic image of papilla discharged from a microsyringe and intradermally injected into a nude mouse by the method of the present invention. A is a papilla labeled with a fluorescent dye DiI in advance, B is a nude mouse showing the transplantation site, C is a tissue observation image (HE staining) of the transplantation site 10 days after intradermal implantation, and D is a tissue image of another transplantation site ( HE staining). E and F are C and D respectively
Nuclear staining images of serial sections of the tissues shown in Figure G and H are the same as DiI.
It is a fluorescent image. The scale bar shows 100 μm.

【図3】この発明の方法によってパピラをマイクロシリ
ンジから排出し、ヒト皮膚に移植した場合の写真像であ
る。Aは皮内移植時所見像、Bは移植後2週目の移植部
位、CとDは移植後6週間における新生毛である。スケー
ルバーは2mmを示す。
FIG. 3 is a photographic image of a papilla discharged from a microsyringe and transplanted to human skin by the method of the present invention. A is the appearance image during intradermal implantation, B is the implantation site 2 weeks after the implantation, and C and D are new hairs 6 weeks after the implantation. Scale bar indicates 2 mm.

───────────────────────────────────────────────────── フロントページの続き (72)発明者 豊島 公栄 広島県東広島市西条中央 6−26−26− 403 (72)発明者 吉里 勝利 広島県東広島市八本松南7−22−13 Fターム(参考) 4C081 AB19 AB20 AC09 BA13 BB04 BC02 CD012 CD34 DA15 DC12 EA02 EA06 4C087 AA01 AA02 BB48 BB63 BB64 CA04 MA01 MA05 NA05 ZA92 4C167 AA71 AA80 BB05 BB06 BB32 CC02 HH14 HH19    ─────────────────────────────────────────────────── ─── Continued front page    (72) Inventor Koei Toyoshima             Hiroshima Prefecture Higashi-Hiroshima City Saijo Chuo 6-26-26-             403 (72) Inventor Yoshiri Victory             7-22-13 Hachimotomatsunami, Higashihiroshima City, Hiroshima Prefecture F-term (reference) 4C081 AB19 AB20 AC09 BA13 BB04                       BC02 CD012 CD34 DA15                       DC12 EA02 EA06                 4C087 AA01 AA02 BB48 BB63 BB64                       CA04 MA01 MA05 NA05 ZA92                 4C167 AA71 AA80 BB05 BB06 BB32                       CC02 HH14 HH19

Claims (3)

【特許請求の範囲】[Claims] 【請求項1】 以下の工程: (1) 液体排出装置の内壁面に高分子多糖溶液を接触さ
せる工程; (2) 微細生体材料を含む液体を排出装置内へ吸入する
工程;および (3) 排出装置より液体を排出する工程、を有すること
を特徴とする微細生体材料の排出方法。
1. The following steps: (1) a step of bringing a high molecular polysaccharide solution into contact with the inner wall surface of the liquid discharge device; (2) a step of sucking a liquid containing a fine biomaterial into the discharge device; and (3) And a step of discharging a liquid from a discharging device.
【請求項2】 微細生体材料が、細胞、組織片または人
工組織片である請求項1の方法。
2. The method according to claim 1, wherein the fine biomaterial is a cell, a piece of tissue or an artificial piece of tissue.
【請求項3】 細胞がパピラ細胞を含む細胞混合物、組
織片がパピラ、人工組織片が培養パピラ細胞から形成し
た人工パピラである請求項2の方法。
3. The method according to claim 2, wherein the cells are a cell mixture containing papilla cells, the tissue pieces are papilla, and the artificial tissue pieces are artificial papilla formed from cultured papilla cells.
JP2002035945A 2002-02-13 2002-02-13 Method for discharging minute organismic material Pending JP2003235990A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2002035945A JP2003235990A (en) 2002-02-13 2002-02-13 Method for discharging minute organismic material

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
JP2002035945A JP2003235990A (en) 2002-02-13 2002-02-13 Method for discharging minute organismic material

Publications (1)

Publication Number Publication Date
JP2003235990A true JP2003235990A (en) 2003-08-26

Family

ID=27777989

Family Applications (1)

Application Number Title Priority Date Filing Date
JP2002035945A Pending JP2003235990A (en) 2002-02-13 2002-02-13 Method for discharging minute organismic material

Country Status (1)

Country Link
JP (1) JP2003235990A (en)

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005053763A1 (en) 2003-12-05 2005-06-16 Biointegrence Inc. Hair growth method
JP2007503876A (en) * 2003-08-26 2007-03-01 アルザ・コーポレーシヨン Devices and methods for intradermal cell transplantation
US7780635B2 (en) 2006-02-09 2010-08-24 Aderans Research Institute, Inc. Apparatus and methods for delivering fluid and material to a subject
WO2010117043A1 (en) * 2009-04-08 2010-10-14 株式会社フェニックスバイオ Method for recovering hair-growing ability of hair follicles, and cell transplantation tool
US7985537B2 (en) 2007-06-12 2011-07-26 Aderans Research Institute, Inc. Methods for determining the hair follicle inductive properties of a composition
USD690004S1 (en) 2012-03-16 2013-09-17 Aderans Research Institute, Inc. Holder for a device for delivering cellular material and physiologic fluids
US9023380B2 (en) 2005-11-22 2015-05-05 Aderans Research Institute, Inc. Hair follicle graft from tissue engineered skin

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2007503876A (en) * 2003-08-26 2007-03-01 アルザ・コーポレーシヨン Devices and methods for intradermal cell transplantation
WO2005053763A1 (en) 2003-12-05 2005-06-16 Biointegrence Inc. Hair growth method
US9023380B2 (en) 2005-11-22 2015-05-05 Aderans Research Institute, Inc. Hair follicle graft from tissue engineered skin
US7780635B2 (en) 2006-02-09 2010-08-24 Aderans Research Institute, Inc. Apparatus and methods for delivering fluid and material to a subject
US8206335B2 (en) 2006-02-09 2012-06-26 Aderans Research Institute, Inc. Apparatus and methods for delivering fluid and material to a subject
US7985537B2 (en) 2007-06-12 2011-07-26 Aderans Research Institute, Inc. Methods for determining the hair follicle inductive properties of a composition
WO2010117043A1 (en) * 2009-04-08 2010-10-14 株式会社フェニックスバイオ Method for recovering hair-growing ability of hair follicles, and cell transplantation tool
USD690004S1 (en) 2012-03-16 2013-09-17 Aderans Research Institute, Inc. Holder for a device for delivering cellular material and physiologic fluids

Similar Documents

Publication Publication Date Title
JP6915224B2 (en) Modified alginates and uses for anti-fibrotic materials
EP2266500B1 (en) Three-dimensional tissue structure
EP1076533B1 (en) Guided development and support of hydrogel-cell compositions
JP3559566B2 (en) Use of autologous dermal fibroblasts for repair of skin and soft tissue defects
JP6970718B2 (en) Compositions and methods for the treatment and repair of tendons
US20070255254A1 (en) Cell delivery system
US20150094808A1 (en) Synthetic scaffolds and organ and tissue transplantation
US20100178683A1 (en) Hair follicle graft from tissue engineered skin
CN104055795B (en) A kind of injectable implant and preparation method thereof
TW200803877A (en) Hair grafts derived from plucked hair
CN105874060A (en) Use of mesothelial cells in tissue bioengineering and artificial tissues
Rogovaya et al. Reconstruction of rabbit urethral epithelium with skin keratinocytes
JP2003235990A (en) Method for discharging minute organismic material
JP4454844B2 (en) Gene therapy medium consisting of dermal sheath tissue
US20170281528A1 (en) Hair Follicles Made Ex Vivo That Can be Inserted into a Recipient for Hair Restoration
BR112013030828B1 (en) PROCESS TO PREPARE A HAIRY LEATHER EQUIVALENT
WO2010117043A1 (en) Method for recovering hair-growing ability of hair follicles, and cell transplantation tool
Li et al. A surgical kit for stem cell-derived retinal pigment epithelium transplants: collection, transportation, and subretinal delivery
CN115177788B (en) PCL composite biological collagen membrane with good mechanical strength and cell activity and application thereof
US20150017131A1 (en) Hair follicles made ex vivo that can be inserted into a recipient for hair restoration
EP4000625A1 (en) Composition for regenerating growth plate
CN117337185A (en) Composition comprising an atelocollagen and Dermal Sheath (DSC) cells, kit for regenerating hair, method for manufacturing a composition for regenerating hair and method for regenerating hair
CN117881437A (en) Collagen IV biological ink
NZ711563B2 (en) Compositions and methods for treating and repairing tendons

Legal Events

Date Code Title Description
A711 Notification of change in applicant

Free format text: JAPANESE INTERMEDIATE CODE: A712

Effective date: 20031031

RD03 Notification of appointment of power of attorney

Free format text: JAPANESE INTERMEDIATE CODE: A7423

Effective date: 20040129

A621 Written request for application examination

Free format text: JAPANESE INTERMEDIATE CODE: A621

Effective date: 20040622

A977 Report on retrieval

Free format text: JAPANESE INTERMEDIATE CODE: A971007

Effective date: 20060929

A131 Notification of reasons for refusal

Free format text: JAPANESE INTERMEDIATE CODE: A131

Effective date: 20061212

A521 Written amendment

Free format text: JAPANESE INTERMEDIATE CODE: A523

Effective date: 20070209

A131 Notification of reasons for refusal

Free format text: JAPANESE INTERMEDIATE CODE: A131

Effective date: 20070403

A521 Written amendment

Free format text: JAPANESE INTERMEDIATE CODE: A523

Effective date: 20070604

A02 Decision of refusal

Free format text: JAPANESE INTERMEDIATE CODE: A02

Effective date: 20070626