JPH11503946A - ヒアルロン酸誘導体を基本とするバイオ適合性物質を支持体として含む人工皮膚 - Google Patents
ヒアルロン酸誘導体を基本とするバイオ適合性物質を支持体として含む人工皮膚Info
- Publication number
- JPH11503946A JPH11503946A JP8532175A JP53217596A JPH11503946A JP H11503946 A JPH11503946 A JP H11503946A JP 8532175 A JP8532175 A JP 8532175A JP 53217596 A JP53217596 A JP 53217596A JP H11503946 A JPH11503946 A JP H11503946A
- Authority
- JP
- Japan
- Prior art keywords
- keratinocytes
- human skin
- fibroblasts
- artificial human
- tissue
- 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.)
- Granted
Links
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Classifications
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- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
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- C12N2502/00—Coculture with; Conditioned medium produced by
- C12N2502/13—Coculture with; Conditioned medium produced by connective tissue cells; generic mesenchyme cells, e.g. so-called "embryonic fibroblasts"
- C12N2502/1323—Adult fibroblasts
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- C12N2503/00—Use of cells in diagnostics
- C12N2503/04—Screening or testing on artificial tissues
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Abstract
Description
Claims (1)
- 【特許請求の範囲】 1.a) 角化細胞が播種され培養された、ヒアルロン酸誘導体を基本とした微小 多孔膜と b) そこで繊維芽細胞が播種され増殖させられた、ヒアルロン酸誘導体を基 本とした下部の不織組織とからなる、人工人間皮膚。 2.繊維芽細胞と角化細胞の間の境界領域に置かれた真皮表皮接合部の蛋白質を 含む、細胞外蛋白質基質より成る請求項1に記載の人工人間皮膚。 3.微小多孔膜と不織組織の両方を調製するのに使用される前記ヒアルロン酸誘 導体がヒアルロン酸エステルであるような請求項1と2の一つに記載の人工人間 皮膚。 4.前記ヒアルロン酸エステルが 75% と 100% の間のエステル化率を持つよう な請求項3に記載の人工人間皮膚。 5.前記エステルがヒアルロン酸ベンジルエステルであるような請求項3と4に 記載の人工人間皮膚。 6.内科及び外科での使用向けの、請求項1-5に記載の人工人間皮膚。 7.深部、II 及び III-度のやけどでの使用、糖尿病性、静脈性、床ずれ性など のグループから選定された多種類の潰瘍への使用、形成外科での使用及びメラニ ン欠損で特徴付けられる疾病への使用のための請求項6に記載の人工人間皮膚。 8.遺伝子異常及び/又は代謝性疾患を矯正する、遺伝子治療に使用するための 請求項6に記載の人工人間皮膚。 9.皮膚と接触して置かれるべく及び化学、薬学、美容整形又は農業の分野で使 用されるべく定められている製品及び/又は物質の可能性のある中毒効果を立証 するための生体外テスト用診断装置としての、請求項1-5の一つに記載の人工 人間皮膚。 10.毛髪移植片のための美容整形の分野で使用する請求項1-5の一つに記載 の人工人間皮膚。 11.生物学的活性化合物、蛋白質、ペプチド及びそれの混合物より成るグルー プから選ばれた少なくとも一つの有効成分を含む薬剤、そこで前記有効成分が不 織組織の繊維及び/又は微小多孔膜に吸着されるような賦形剤として、請求項1 -5の一つに記載の人工人間皮膚を含む制御された放出薬剤。 12.前記蛋白質が、繊維芽細胞成長因子、形質転換成長因子β、角化細胞成長 因子、神経成長因子、又はフィブロネクチン、ラミニン、コラーゲン、又は凝固 カスケードに含まれる蛋白質より成るグループから選ばれた細胞外基質の蛋白質 より成るグループから選ばれた、成長因子であるような請求項11に記載の制御 された放出薬剤。 13.前記細胞外基質の蛋白質が細胞外基質の繊維に吸着されているような請求 項12に記載の制御された放出薬剤。 14.以下の段階より成る請求項1-5の一つに記載の人工人間皮膚を調製する 方法: i) ヒアルロン酸誘導体を基本にした微小多孔膜上に、1,000 から 100,000 細胞 /cm2までの密度範囲で培養する角化細胞の播種及び、部分的な又は完全な合流 が達成するまで、子牛の血清の使用を含む従来の技術に従った前記膜上の前記Kc の拡張工程、 ii) 10% 子牛の血清を含んでいる DMEM 中で通常の技術により真皮から又は他の 部位から分離した繊維芽細胞の培養、プラスチックの上に播種して増殖するため のこれら繊維芽細胞の条件付け、不織組織へのその繊維芽細胞の播種及び(得ら れた人工皮膚が保存されねばならない場合の)凍結まで又は移植までこの培養を 続ける工程、 iii) 微小多孔膜の外側の縁が皿の底の触れないように又それが下に横たわる不 織組織を越えて僅かに拡がるように注意しながら、繊維芽細胞によりコロニー化 している不織組織の上に既に形成された上皮層を横たえる工程、 iv) 微小多孔膜上の上皮細胞層を繊維芽細胞によりコロニー化している下に横た わる不織組織に、コラーゲン、フィブリン、フィブリン膠より成るグループから 選ばれた生物学的接着剤により、一緒に任意選択的に固着する、 v) 段階i)に使用したのと同じ培地を十分な量加える、微小多孔膜の上部層の角 化細胞が空気-液体の境界にあるように浸された培養物を保持する、最初と続く 培地の変化で、1μg/ml に等しい濃度のアスコルビン酸を添加、移植又は凍結 まで培養を続ける工程。 15.段階i)において、角化細胞の播種密度が 5,000 と 25,000 細胞/cm2の 間より成り、又細胞は非増殖の 3T3-ねずみの繊維芽細胞の“フィーダー層”の 存在下、CEC 培地中で増殖するような請求項14に記載の方法。 16.以下の段階より成る請求項1-5の一つに記載の人工人間皮膚を調製する 方法: i′)不織組織に繊維芽細胞を播種して凍結又は移植まで培養を続ける工程、 ii′)不織組織の上に微小多孔膜を、その縁が下に横たわる組織を僅かに越えて 拡がるように横たえる、又従来の方法で用いられる生物学的接着剤又は機械的手 段により前記膜を固着する工程、 iii′)微小多孔膜上へ角化細胞の播種、プラスチック皿の上で角化細胞の成長 に適した上記”化学的に定義された”培地を用いて合流を達成するまで前記角化 細 胞の増殖工程、 iv′)前の段階で用いたのと同じ培地の十分な量を加えて、培養物は微小多孔膜 の上部層の角化細胞が空気-液体境界にあるように浸される、最初と続く培地の 変化で、1μg/mlに等しい濃度のアスコルビン酸を添加し、移植まで培養を続け る工程。 17.皮膚が保存されねばならない時、更に凍結保存剤の存在で前記人工皮膚を 凍結保存する段階を構成するような、請求項14と15の一つに記載の工程、又 は請求項16に記載の工程。 18.繊維芽細胞と角化細胞が一次培養物から又は凍結した在庫品から出て来る ような、請求項14と15の一つに記載の工程、又は請求項16に記載の工程。
Applications Claiming Priority (3)
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IT95A000083 | 1995-03-08 | ||
IT95PD000083A IT1281870B1 (it) | 1995-04-27 | 1995-04-27 | Pelle artificiale umana costituita da materiali biocompatibili a base di derivati dell'acido ialuronico |
PCT/EP1996/001734 WO1996033750A1 (en) | 1995-04-27 | 1996-04-25 | Artificial skin containing as support biocompatible materials based on hyaluronic acid derivatives |
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JPH11503946A true JPH11503946A (ja) | 1999-04-06 |
JP3646882B2 JP3646882B2 (ja) | 2005-05-11 |
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JP53217596A Expired - Fee Related JP3646882B2 (ja) | 1995-04-27 | 1996-04-25 | ヒアルロン酸誘導体を基本とするバイオ適合性物質を支持体として含む人工皮膚 |
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US (1) | US6110208A (ja) |
EP (1) | EP0822839B1 (ja) |
JP (1) | JP3646882B2 (ja) |
AT (1) | ATE206058T1 (ja) |
AU (1) | AU700762B2 (ja) |
CA (1) | CA2219272A1 (ja) |
DE (1) | DE69615549T2 (ja) |
DK (1) | DK0822839T3 (ja) |
ES (1) | ES2162052T3 (ja) |
IT (1) | IT1281870B1 (ja) |
PT (1) | PT822839E (ja) |
WO (1) | WO1996033750A1 (ja) |
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1995
- 1995-04-27 IT IT95PD000083A patent/IT1281870B1/it active IP Right Grant
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1996
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- 1996-04-25 JP JP53217596A patent/JP3646882B2/ja not_active Expired - Fee Related
- 1996-04-25 ES ES96913526T patent/ES2162052T3/es not_active Expired - Lifetime
- 1996-04-25 EP EP96913526A patent/EP0822839B1/en not_active Expired - Lifetime
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- 1996-04-25 CA CA002219272A patent/CA2219272A1/en not_active Abandoned
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JP2018513724A (ja) * | 2015-03-26 | 2018-05-31 | ユニベルシテ ドゥ ボルドー | 皮膚再建の方法 |
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US6110208A (en) | 2000-08-29 |
PT822839E (pt) | 2002-03-28 |
ATE206058T1 (de) | 2001-10-15 |
EP0822839B1 (en) | 2001-09-26 |
ITPD950083A0 (it) | 1995-04-27 |
WO1996033750A1 (en) | 1996-10-31 |
DK0822839T3 (da) | 2002-01-21 |
ITPD950083A1 (it) | 1996-10-27 |
DE69615549D1 (de) | 2001-10-31 |
CA2219272A1 (en) | 1996-10-31 |
EP0822839A1 (en) | 1998-02-11 |
DE69615549T2 (de) | 2002-06-06 |
JP3646882B2 (ja) | 2005-05-11 |
ES2162052T3 (es) | 2001-12-16 |
AU5647996A (en) | 1996-11-18 |
AU700762B2 (en) | 1999-01-14 |
IT1281870B1 (it) | 1998-03-03 |
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