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Method of forming normal regenerated tissue, the normal regenerated tissue, and method of calibrating senstivity and so on

外国特許コード F110004185
整理番号 Y0223WO
掲載日 2011年7月12日
出願国 欧州特許庁(EPO)
出願番号 02777835
公報番号 1444994
公報番号 1444994
出願日 平成14年10月15日(2002.10.15)
公報発行日 平成16年8月11日(2004.8.11)
公報発行日 平成26年7月30日(2014.7.30)
国際出願番号 JP2002010674
国際公開番号 WO2003039611
国際出願日 平成14年10月15日(2002.10.15)
国際公開日 平成15年5月15日(2003.5.15)
優先権データ
  • 2002JP010674 (2002.10.15) WO
  • 特願2001-317502 (2001.10.15) JP
発明の名称 (英語) Method of forming normal regenerated tissue, the normal regenerated tissue, and method of calibrating senstivity and so on
発明の概要(英語) A normal regenerated tissue is formed by exposing to radiation a connective tissue or a supporting tissue originating in an organ to thereby form a feeder layer and then transplanting epithelial cells thereon to form a stratified structure.
By conveniently and surely providing regenerated tissue by the 3-dimensional culture with the use of a human-origin normal tissue as a base, it is possible to construct systems for assessing effects and side effects of chemicals such as drugs or assessing sensitivities thereof with the use of regenerated tissues as models of corresponding tissues respectively. <IMAGE>
特許請求の範囲(英語) [claim1]
1. A method for forming a normal regenerated tissue comprising irradiating an organ-derived connective tissue or its constituent cells or supporting tissue to form a feeder layer followed by transplanting epithelial cells to form a stratified structure of the epithelial cells, wherein the connective tissue or its constituent cells or supporting tissue and the epithelial tissue are orthotopic with regard to the originating organ.
[claim2]
2. The method for forming a normal regenerated tissue according to Claim 1 wherein the epithelial cells derive from nerve, oral mucosa, bronchus, mammary gland, liver, prostate or kidney.
[claim3]
3. The method for forming a normal regenerated tissue according to Claim 1 or Claim 2 wherein the connective tissue or its constituent cells or supporting tissue are at least any of the organ-derived fibroblasts, endothelial cells or its constituent tissue.
[claim4]
4. The method for forming a normal regenerated tissue according to any one of Claims 1 to 3 wherein vascular endothelial cells are transplanted on organ-derived fibroblasts and then an irradiation is effected to form a feeder layer, and then the epithelial cells are transplanted to form a stratified structure.
[claim5]
5. The method for forming a normal regenerated tissue according to any one of Claims 1 to 4 wherein after transplanting the epithelial cells at least one extracellular matrix is added to form the epithelial cells in a stratified structure.
[claim6]
6. The method for forming a normal regenerated tissue according to Claim 5 wherein the extracellular matrix is a structural component or adhesion molecule of an extracellular substrate.
[claim7]
7. The method for forming a normal regenerated tissue according to Claim 5 or 6 wherein the extracellular matrix is at least one of collagen, elastin, proteoglycan, fibronectin, laminin and tenascin.
[claim8]
8. The method for forming a normal regenerated tissue according to Claim 7 wherein after transplanting a collagen or collagen with fibronectin as well as laminin are added to form the epithelial cells in a stratified structure.
[claim9]
9. The method for forming a normal regenerated tissue according to any one of Claims 1 to 8 wherein the connective tissue or its constituent cells or supporting tissue is co-cultured with at least one of heterogenous constituent cells derived from an orthotopic organ or a culture supernatant thereof and thereafter irradiated to form a feeder layer.
[claim10]
10. The method for forming a normal regenerated tissue according to any one of Claims 1 to 9 wherein heterogenous constituent cells deriving from an orthotopic organ, epithelial cells and a culture supernatant thereof is co-cultured to form a stratified structure.
[claim11]
11. The method for forming a normal regenerated tissue according to any one of Claims 1 to 10 wherein the irradiation is an X-ray or gamma-ray irradiation.
[claim12]
12. The method for forming a normal regenerated tissue according to any one of Claims 1 to 11 wherein the stratified structure of the epithelial cells is formed by a culture at a carbon dioxide gas concentration of 5 to 15%, air concentration of 85 to 95% at a culture temperature of 20 to 40 deg.C.
[claim13]
13. A normal regenerated tissue comprising a stratified structure of epithelial cells on an irradiated feeder layer from an organ-derived connective tissue or constituent cells or supporting tissue, wherein the connective tissue or its constituent cells or supporting tissue and the epithelial tissue are orthotopic with regard to the deriving organ.
[claim14]
14. The normal regenerated tissue according to Claim 13 wherein the epithelial cells derive from nerve, oral mucosa, bronchus, mammary gland, liver, prostate or kidney.
[claim15]
15. The normal regenerated tissue according to Claim 13 or 14 wherein the connective tissue or its constituent cell or supporting tissue are at least any of the organ-derived fibroblasts, endothelial cells or its constituent tissue.
[claim16]
16. A normal regenerated tissue comprising an irradiated feeder layer consisting of a lower layer of organ-derived fibroblasts and an upper layer of vascular endothelial cells and epithelial cells in a stratified structure placed on the feeder layer, wherein the fibroblasts and the vascular endothelial cells and epithelial cells are orthotopic with regard to the deriving organ.
[claim17]
17. The normal regenerated tissue according to claim 16 wherein the epithelial cells derive from nerve, oral mucosa, bronchus, mammary gland, liver, prostate or kidney.
[claim18]
18. The normal regenerated tissue according to any one of Claims 13 to 17 wherein the epithelial cells are in a stratified structure consisting of four or more layers.
[claim19]
19. The normal regenerated tissue according to any one of Claims 13 to 18 additionally comprising a basal plate.
[claim20]
20. The normal regenerated tissue which constitutes a regenerated organ model body of a normal regenerated tissue according to any one of Claims 13 to 19.
[claim21]
21. A method for assessing a sensitivity of cancer cells comprising inhibiting the proliferation of regenerated epithelial cells by irradiating a normal regenerated tissue according to any one of Claims 13 to 20 followed by transplanting the cancer cells on the epithelial cells in a stratified structure followed by adding a chemical substance or irradiation.
[claim22]
22. The method according to Claim 21 wherein the assessment is effected at a low oxygen concentration.
[claim23]
23. The method according to Claim 21 or 22 comprising supplementing at least one of collagen or other extracellular matrixes after transplanting cancer cells or transplanting cancer cells after supplementing at least one of collagen or other extracellular matrixes, followed by assessing the sensitivity.
[claim24]
24. A method for assessing an angiogenetic ability comprising inhibiting the proliferation of regenerated epithelial cells by irradiating a normal regenerated tissue according to any one of Claims 13 to 20 followed by transplanting the cancer cells on the epithelial cells in a stratified structure followed by supplementing at least one of collagen or other extracellular matrixes on which then vascular endothelial cells are transplanted followed by assessing the angiogenetic ability of the cancer cells in response to the addition of a chemical substance.
[claim25]
25. A method for assessing an angiogenetic ability comprising inhibiting the proliferation of regenerated epithelial cells by irradiating a normal regenerated tissue according to any one of Claims 13 to 20 followed by transplanting the cancer cells on the epithelial cells in a stratified structure followed by mounting at least one of collagen or other extracellular matrixes followed by inverting the entire structure whereby assessing a motility or invasion ability.
[claim26]
26. A method for assessing a sensitivity of a normal regenerated tissue according to any one of Claims 13 to 20 comprising an exposure of the tissue to a chemical substance or irradiation.
[claim27]
27. A method for assessing a gene transduction comprising assessing the efficiency of the gene transduction in a normal regenerated tissue according to any one of Claims 13 to 20.
[claim28]
28. The method according to any one of Claims 21 to 27 wherein the irradiation for inhibiting the proliferation of regenerated epithelial cells is an X-ray or gamma-ray irradiation.
  • 出願人(英語)
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
  • 発明者(英語)
  • NAKATSUGAWA SHIGEKAZU
国際特許分類(IPC)
欧州特許分類/主・副
  • A61L027/36B
  • A61L027/38B4
  • C12N005/06T
  • K61K035/12
  • M12N502/08P
  • M12N502/09M
  • M12N502/24M
  • M12N502/25K
  • M12N502/28
  • M12N502/30
  • M12N503/00
  • M12N503/04
指定国 Contracting States: AT BE BG CH CY CZ DE DK EE ES FI FR GB GR IE IT LI LU MC NL PT SE SK TR
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