骨髄細胞を用いた新しい皮膚再生医療の開発

• Project/Area Number: 15790563
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Dermatology
• Research Institution: Hokkaido University
• Principal Investigator: 阿部 理一郎 北海道大学, 病院, 助手 (60344511)
• Project Period (FY): 2003 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥3,600,000 (Direct Cost: ¥3,600,000); Fiscal Year 2004: ¥1,400,000 (Direct Cost: ¥1,400,000); Fiscal Year 2003: ¥2,200,000 (Direct Cost: ¥2,200,000)
• Keywords: 再生医療 / 組織間幹細胞 / 表皮細胞 / 骨髄細胞 / 人工真皮

Research Abstract

近年の再生医学の進歩により、骨髄由来の細胞は表皮細胞をはじめとする皮膚構成細胞に分化しうることが明らかになった。しかしながら骨髄単核球が皮膚構成細胞の大部分に分化が可能であるにも関わらず、実際in-vitroで生理的に機能しうる皮膚を再生させることは現在においても不可能である。本研究の目的は、骨髄細胞を用いた皮膚再生医療の開発を行い、実際臨床応用の手がかりを得ることである。
1.皮膚再生に必要なそれぞれの細胞(表皮細胞、線維芽細胞、内皮細胞等)におけるニッチの解析。
(1)酸素濃度、温度、培養液等の培養条件。
(2)添加する液性因子(EGF、bFGF、TGFβなどの増殖因子、サイトカイン等)。
(3)添加、または培養フラスコにコートする接着分子(コラーゲン等)。
(4)共培養細胞(表皮細胞、線維芽細胞、内皮細胞、脂肪細胞等)。
(5)人工真皮(生体吸収性高分子)
以上の条件を変え、または組み合わせて培養を行い、表皮細胞との共培養の条件が最も皮膚細胞に分化することを見出した。
2.動物モデルを用いた、生体内における機能的な皮膚臓器の再生の検討。
(1)免疫不全マウスに皮膚欠損創を作成し、ヒト骨髄単核球を含んだ人工真皮を貼付することで、人工真皮中の骨髄単核球が線維芽細胞、内皮細胞に分化することを確認した。
(2)GFPトランスゲニックマウスの骨髄を正常マウスに移植したキメラマウスにおいて皮膚欠損創を作成したところ、骨髄由来の表皮細胞、ならびに線維芽細胞、内皮細胞に分化することを確認した。

Research Products

• [Journal Article] Overexpression of pigment epithelium-derived factor decreases angiogenesis and inhibits the growth of human malignant melanoma cells in vivo. (2004)
  • Author(s): Abe R., et al.
  • Journal Title: Am J Pathol 164 Pages: 1225-1232
• [Journal Article] Regulation of human malignant melanoma growth and metastasis by AGE-AGE receptor interactions. (2004)
  • Author(s): Abe R., et al.
  • Journal Title: J Invest Dermatol 122 Pages: 461-467
• [Journal Article] Inhibition by minodronate of vascular endothelial growth factor signaling suppresses melanoma growth in nude mice. (2004)
  • Author(s): Yamagishi S., et al.
  • Journal Title: Am J Pathol 165 Pages: 1865-1874
• [Journal Article] Macrophage migration inhibitory factor (MIF) is induced by thrombin and factor Xa in endothelial cells. (2004)
  • Author(s): Shimizu T., et al.
  • Journal Title: J Biol Chem 279 Pages: 13729-13737
• [Journal Article] Expression of maf in the skin during rat embryonic development. (2004)
  • Author(s): Ogata A., et al.
  • Journal Title: Acta Histochem 106 Pages: 65-67
• [Journal Article] Macrophage Migration Inhibitory Factor (MIF) mediates the up-regulation of matrix metalloproteinase-1 by the stimulation of ultraviolet A irradiation in human dermal fibroblasts. (2004)
  • Author(s): Watanabe H., et al.
  • Journal Title: J Biol Chem 279 Pages: 1676-1683
• [Journal Article] Cetirizine, an H1-receptor antagonist, suppresses the expression of macrophage migration inhibitory factor (MIF) : its potential anti-inflammatory action. (2004)
  • Author(s): Shimizu T., et al.
  • Journal Title: Clin Exp Allergy 34 Pages: 103-110
• [Journal Article] Direct injection of plasmid DNA into the skin induces dermatitis by activation of monocytes through toll-like receptor 9.
  • Author(s): Sawamura D., et al.
  • Journal Title: J Gene Med (In press)
• [Journal Article] Promoter region polymorphism of macrophage migration inhibitory factor (MIF) is strong risk factor for young onset of extensive alopecia areata.
  • Author(s): Shimizu T., et al.
  • Journal Title: Genes Immun (In press)
• [Publications] Abe R., et al.: "Overexpression of pigment epithelium-derived factor decreases angiogenesis and inhibits the growth of human malignant melanoma cells in vivo."Am J Pathol. 164. 1225-1232 (2004)
• [Publications] Abe R., et al.: "Regulation of human malignant melanoma growth and metastasis by AGE-AGE receptor interactions."J Invest Dermatol. 122. 461-467 (2004)
• [Publications] Abe R., et al.: "Toxic epidermal necrolysis and Stevens-Johnson syndrome are induced by soluble Fas ligand."Am J Pathol. 162. 1515-1520 (2003)
• [Publications] Shimizu T., et al.: "Macrophage migration inhibitory factor (MIF) is induced by thrombin and factor Xa in endothelial cells."J Biol Chem. (In press).
• [Publications] Ogata A., et al.: "Expression of maf in the skin during rat embryonic development."Acta Histoehem. (In press).
• [Publications] Watanabe H., et al.: "Macrophage Migration Inhibitory Factor (MIF) mediates the up-regulation of matrix metalloproteinase-1 by the stimulation of ultraviolet A irradiation in human dermal fibroblasts."J Biol Chem. 279. 1676-1683 (2004)
• [Publications] Shimizu T., et al.: "Impaired contact hypersensitivity in macrophage migration inhibitory factor (MIF) - deficient mice."Eur J Immunol. 33. 1478-1487 (2003)
• [Publications] Shimizu T., et al.: "Cetirizine, an H1-receptor antagonist, suppresses the expression of macrophage migration inhibitory factor (MIF):its potential anti-inflammatory action."Clin Exp Allergy. 34. 103-110 (2004)
• [Publications] Yamagishi S., et al.: "Nifedipine inhibits apoptotic cell death of cultured endothelial cells induced by tumor necrosis factor-α."Durg Exp Clin Res. 29. 141-145 (2003)