JP4550582B2 - 毛髪誘導細胞の培養 - Google Patents
毛髪誘導細胞の培養 Download PDFInfo
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- JP4550582B2 JP4550582B2 JP2004550840A JP2004550840A JP4550582B2 JP 4550582 B2 JP4550582 B2 JP 4550582B2 JP 2004550840 A JP2004550840 A JP 2004550840A JP 2004550840 A JP2004550840 A JP 2004550840A JP 4550582 B2 JP4550582 B2 JP 4550582B2
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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
- C12N5/0625—Epidermal cells, skin cells; Cells of the oral mucosa
- C12N5/0627—Hair cells
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- 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
- A61K8/985—Skin or skin outgrowth, e.g. hair, nails
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- 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
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- 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
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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
- C12N2502/00—Coculture with; Conditioned medium produced by
- C12N2502/24—Genital tract cells, non-germinal cells from gonads
- C12N2502/246—Cells of the male genital tract, non-germinal testis cells
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- Dermatology (AREA)
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- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Description
本発明は更に、男性型はげ頭症の処置(例えば化粧法)のための、本明細書に記載の方法を用いて得られる例えばDP細胞及び/又はDS細胞等の培養毛髪誘導細胞の使用を提供する。
調整培地(CM)の調製
CM試験用の増殖細胞がコンフルエント段階に近づいた時、新鮮培地を供給した。5日後、培地を除き、0.22μmフィルターでろ過し、細胞及び残骸を除いた。当該CMは、新鮮培地と1:1(体積:体積)の割合で混合し、培養した真皮乳頭細胞を供給するために用いた。CMは標準的条件下で増殖したヒトのケラチノサイトから生成させた。
以下の外胚葉性起源の細胞種を試験した:2種のヒト乳癌細胞株;MCF−7(American Type Culture Collection; ATCC)及びMDA−MB−231(ATCC)及び胎児性腫瘍細胞株であるN−テラ−2(P. Andrews, Sheffield, UK)、ヒト乳房上皮細胞(Cascade Biologics, Oregon, USA)及びヒトメラノサイト(Cambrex, New Jersey, USA)。以下の中胚葉起源の細胞種を試験した:ヒト真皮繊維芽細胞(HDF、新鮮な摘出ヒト組織から単離し、標準的条件下で増殖した)、骨格筋芽細胞(Cambrex)、前立腺上皮細胞(Cambrex)。試験に使用できる他の細胞種は:腎上皮細胞、内皮細胞及び免疫系細胞(中胚葉起源);及び肝細胞及び膀胱上皮組織(外胚葉起源)。特に断らなければ、細胞種は供給者によって推奨される条件下で培養する。
ヒトDPは、毛髪リンパ小節からDPを顕微解剖した後、培地中に入れた。標準培地で5日後、培地を試験CMと交換した。培養液を2〜3日毎に交換し、培養をコンフルエントになるまで継代した。培養を当該方法で数継代維持した。図2及び3に示す細胞は、3継代での写真である。
皮膚再構成試験(Lichti, et al., 1993, J, Invest. Dermatol. 101: 124S-129S; and Weinberg et al., 1993, J Invest. Dertmatol. 100: 229-236)を培養毛髪誘導(例えばDP)細胞による毛髪誘導を試験するために使用した。麻酔したマススの背中に円形の十分に深い傷を作り、移植チェンバー(graft chamber)をその傷中に設置した。培養真皮乳頭細胞又は真皮繊維芽細胞(負のコントロールとして)と混合した齧歯動物のケラチノサイトを、移植チェンバーを通して注射した。1週間後、チェンバーを外し、外傷に包帯をした。移植細胞は十分に厚い皮膚を形成し、毛髪誘導真皮乳頭細胞が存在している場合には新しい毛髪リンパ小節も形成された。DP細胞によって誘導された新しい毛髪が移植から4週間以内に現れた。
真皮乳頭細胞は、最初に単離された時に特徴的な形態を有する。当該細胞は小さく、丸く又は多角形で、かつ無規律な群集中で増殖する(図1)。いくつかの初期の群集の中には、分割していない大きな平坦な細胞が存在する。標準的培地中での培養で時間超過すると、大きな平坦な細胞の割合が増加する。当該細胞は毛髪リンパ小節形成を誘導する能力が低い。従って、理想的な培養条件下で観察された形態は、毛髪誘導能の最初の兆候として見ることができる。
Claims (21)
- 前立腺上皮細胞、及び前立腺上皮細胞によって調整された培地を含む培養液中で真皮乳頭(DP)細胞を培養する工程を含む、DP細胞の培養方法。
- 前記DP細胞の毛髪誘導能が維持されている、請求項1記載の方法。
- 前記調整培地が前立腺上皮細胞の細胞株を使用して得られる、請求項1又は2記載の方法。
- 前記細胞株が樹立された細胞株である、請求項3記載の方法。
- 前記培養液が組み換え遺伝子及び/又はその組み換え産物を含まない、請求項1〜4のいずれか1項記載の方法。
- 前記培養液がウイルス性ベクターを含まない、請求項1〜5のいずれか1項記載の方法。
- 前記調整培地が使用前に凍結されている、請求項1〜6のいずれか1項記載の方法。
- 前記調整培地が、10μg/mlを超える総蛋白含量を有する無血清成分を含む、請求項1〜7のいずれか1記載の方法。
- 前記調整培地が、100μg/mlを超える総蛋白含量を有する無血清成分を含む、請求項1〜8のいずれか1記載の方法。
- 前記調整培地が、1mg/mlを超える総蛋白含量を有する無血清成分を含む、請求項1〜8のいずれか1記載の方法。
- 前記調整培地が使用前に濃縮されている、請求項1〜10のいずれか1項記載の方法。
- 前記調整培地が使用前に限外濾過によって濃縮されている、請求項1〜11のいずれか1項記載の方法。
- 3以上の継代のための培養液中で前記DP細胞を継代培養する工程を更に含む、請求項1〜11のいずれか1項記載の方法。
- 7以上の継代のための培養液中で前記DP細胞を継代培養する工程を更に含む、請求項1〜12のいずれか1項記載の方法。
- 培養もしくは継代培養されたDP細胞を採取又は単離する工程を更に含む、請求項1〜14のいずれか1項記載の方法。
- 前記DP細胞が非表皮性組織に対して同種異系である、請求項1〜15のいずれか1項記載の方法。
- 前記DP細胞が非表皮性組織に対して自系である、請求項1〜16のいずれか1項記載の方法。
- 哺乳動物種の真皮乳頭(DP)細胞の長期間培養方法であって、前立腺上皮細胞、及び前立腺上皮細胞よって調整された培地を含む細胞培養液中でDP細胞を培養かつ継代培養し、それによってDP細胞を増殖すると同時にそれらの毛髪誘導能を維持する工程を含む、方法。
- 移植のための真皮乳頭(DP)細胞を提供しかつ維持するための方法であって、対象から得られたDP細胞を、請求項1〜18のいずれか1項記載の条件下で培養する工程を含む、方法。
- 真皮乳頭(DP)細胞、並びに
前立腺上皮細胞、及び前立腺上皮細胞によって調整された培地を含む培養液、
を含む組成物。 - 真皮乳頭(DP)細胞の培養のための培養液であって、前立腺上皮細胞、及び前立腺上皮細胞によって調整された培地を含み、かつ該DP細胞の毛髪誘導能を維持することができる、培養液。
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US42611102P | 2002-11-14 | 2002-11-14 | |
PCT/GB2003/004949 WO2004044188A1 (en) | 2002-11-14 | 2003-11-14 | Cultivation of hair inductive cells |
Publications (3)
Publication Number | Publication Date |
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JP2006506063A JP2006506063A (ja) | 2006-02-23 |
JP2006506063A5 JP2006506063A5 (ja) | 2007-01-25 |
JP4550582B2 true JP4550582B2 (ja) | 2010-09-22 |
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ID=32313110
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
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JP2004550840A Expired - Fee Related JP4550582B2 (ja) | 2002-11-14 | 2003-11-14 | 毛髪誘導細胞の培養 |
Country Status (8)
Country | Link |
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US (1) | US7785876B2 (ja) |
EP (1) | EP1563061B1 (ja) |
JP (1) | JP4550582B2 (ja) |
AU (1) | AU2003282244B2 (ja) |
CA (2) | CA2505409C (ja) |
DK (1) | DK1563061T3 (ja) |
ES (1) | ES2387617T3 (ja) |
WO (1) | WO2004044188A1 (ja) |
Families Citing this family (20)
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US20030161815A1 (en) * | 2002-02-12 | 2003-08-28 | Intercytex Limited | Cell delivery system |
WO2004044188A1 (en) | 2002-11-14 | 2004-05-27 | Intercytex Limited | Cultivation of hair inductive cells |
US20050233450A1 (en) * | 2004-01-12 | 2005-10-20 | Goetinck Paul F | Methods of inducing hair growth |
US20060154365A1 (en) * | 2004-08-30 | 2006-07-13 | Anthony Ratcliffe | Cultured three dimensional tissues and uses thereof |
KR100616752B1 (ko) * | 2004-11-29 | 2006-08-31 | 박정극 | 모낭 유도 능력이 있는 모유두 조직의 제조방법 |
US20070087437A1 (en) * | 2005-10-14 | 2007-04-19 | Jifan Hu | Methods for rejuvenating cells in vitro and in vivo |
AU2006304516A1 (en) * | 2005-10-17 | 2007-04-26 | Aderans Research Institute, Inc. | Method of delivering hair follicle progenitor cells to the skin |
TW200800240A (en) | 2005-11-22 | 2008-01-01 | Aderans Res Inst Inc | Hair follicle graft from tissue engineered skin |
US9109204B2 (en) | 2006-02-28 | 2015-08-18 | The Trustees Of Columbia University In The City Of New York | Methods for compact aggregation of dermal cells |
GB0605450D0 (en) | 2006-03-17 | 2006-04-26 | Intercytex Ltd | Cell co-culture |
KR100771171B1 (ko) * | 2006-07-05 | 2007-10-29 | 재단법인서울대학교산학협력재단 | 모낭 줄기세포의 분리, 증식 및 분화 방법 및 대머리치료용 조성물 |
US7922688B2 (en) | 2007-01-08 | 2011-04-12 | Restoration Robotics, Inc. | Automated delivery of a therapeutic or cosmetic substance to cutaneous, subcutaneous and intramuscular tissue regions |
US7985537B2 (en) | 2007-06-12 | 2011-07-26 | Aderans Research Institute, Inc. | Methods for determining the hair follicle inductive properties of a composition |
WO2009089380A2 (en) * | 2008-01-08 | 2009-07-16 | The Trustees Of Columbia University In The City Of New York | Methods for p2ry5 mediated regulation of hair growth and mutants thereof |
GB0916370D0 (en) * | 2009-09-18 | 2009-10-28 | Avecia Biolog Ltd | Compositions |
US9198911B2 (en) | 2010-11-02 | 2015-12-01 | The Trustees Of Columbia University In The City Of New York | Methods for treating hair loss disorders |
KR102027394B1 (ko) | 2010-11-02 | 2019-10-01 | 더 트러스티스 오브 콜롬비아 유니버시티 인 더 시티 오브 뉴욕 | 탈모 질환의 치료 방법 |
CN104334191A (zh) | 2012-03-29 | 2015-02-04 | 纽约市哥伦比亚大学托管会 | 治疗毛发脱落疾病的方法 |
FR3072972B1 (fr) * | 2017-10-30 | 2023-03-10 | Oreal | Procede de preparation in vitro d’equivalents de papille dermique et de follicule pileux |
GB201816902D0 (en) * | 2018-10-17 | 2018-11-28 | Hairclone Ltd | Hair Rejuvenation |
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GB8604360D0 (en) | 1986-02-21 | 1986-03-26 | Univ Dundee | Stimulation of hair growth |
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DE10005329A1 (de) | 2000-02-08 | 2001-09-20 | Albert Scheller | Humanisiertes, vitalisiertes Haarbewachsen und Haarwiederbewachsen mit subkutanen Kanüleneinstich-Haut-Färbungs-Zeichnungsverfahren |
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WO2004044188A1 (en) | 2002-11-14 | 2004-05-27 | Intercytex Limited | Cultivation of hair inductive cells |
-
2003
- 2003-11-14 WO PCT/GB2003/004949 patent/WO2004044188A1/en active Application Filing
- 2003-11-14 DK DK03773862.2T patent/DK1563061T3/da active
- 2003-11-14 US US10/534,428 patent/US7785876B2/en not_active Expired - Lifetime
- 2003-11-14 CA CA2505409A patent/CA2505409C/en not_active Expired - Lifetime
- 2003-11-14 AU AU2003282244A patent/AU2003282244B2/en not_active Expired
- 2003-11-14 CA CA2958146A patent/CA2958146C/en not_active Expired - Lifetime
- 2003-11-14 EP EP03773862A patent/EP1563061B1/en not_active Expired - Lifetime
- 2003-11-14 ES ES03773862T patent/ES2387617T3/es not_active Expired - Lifetime
- 2003-11-14 JP JP2004550840A patent/JP4550582B2/ja not_active Expired - Fee Related
Also Published As
Publication number | Publication date |
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US20050272150A1 (en) | 2005-12-08 |
JP2006506063A (ja) | 2006-02-23 |
EP1563061A1 (en) | 2005-08-17 |
WO2004044188A1 (en) | 2004-05-27 |
AU2003282244A1 (en) | 2004-06-03 |
DK1563061T3 (da) | 2012-10-01 |
EP1563061B1 (en) | 2012-06-20 |
CA2505409C (en) | 2017-04-04 |
CA2505409A1 (en) | 2004-05-27 |
AU2003282244B2 (en) | 2008-11-20 |
US7785876B2 (en) | 2010-08-31 |
ES2387617T3 (es) | 2012-09-27 |
CA2958146C (en) | 2019-08-20 |
CA2958146A1 (en) | 2004-05-27 |
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