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Method for isolating smooth muscle stem cells

外国特許コード F120006109
整理番号 S2008-0528
掲載日 2012年1月6日
出願国 アメリカ合衆国
出願番号 93730509
公報番号 20110033428
公報番号 8747838
出願日 平成21年4月13日(2009.4.13)
公報発行日 平成23年2月10日(2011.2.10)
公報発行日 平成26年6月10日(2014.6.10)
国際出願番号 JP2009057809
国際公開番号 WO2009125877
国際出願日 平成21年4月13日(2009.4.13)
国際公開日 平成21年10月15日(2009.10.15)
優先権データ
  • 特願2008-103867 (2008.4.11) JP
  • 2009JP057809 (2009.4.13) WO
発明の名称 (英語) Method for isolating smooth muscle stem cells
発明の概要(英語) This invention relates to a method for isolating smooth muscle stem cells derived from mammalian smooth muscle comprising bringing mammalian smooth muscle cells into contact with a fluorescence-labeled anti-CD45 antibody, anti-CD34 antibody, and anti-CD49f antibody, and isolating cells that would not bind to the anti-CD45 antibody but would bind to the anti-CD34 antibody and the anti-CD49f antibody.
特許請求の範囲(英語) [claim1]
1. A method for isolating a population of uterine smooth muscle-derived stem cells from mammalian uterine smooth muscle, the method comprising: bringing the mammalian uterine smooth muscle cells dispersed from uterine smooth muscle into contact with an anti-CD34 antibody and an anti-CD49f antibody labeled with fluorescent dyes; and
isolating a population of uterine smooth muscle-derived stem cells that bind to the anti-CD34 antibody and the anti-CD49f antibody.
[claim2]
2. A method for isolating a population of uterine smooth muscle-derived stem cells from mammalian uterine smooth muscle, the method comprising: bringing the mammalian uterine smooth muscle cells into contact with an anti-CD45 antibody, an anti-CD34 antibody, and an anti-CD49f antibody labeled with fluorescent dyes; and
isolating a population of uterine smooth muscle-derived stem cells that do not bind to the anti-CD45 antibody but bind to the anti-CD34 antibody and the anti-CD49f antibody.
[claim3]
3. The method according to claim 1, further comprising sorting the mammalian uterine smooth muscle cells via flow cytometry.
[claim4]
4. A method for isolating a population of uterine smooth muscle-derived stem cells from mammalian smooth muscle, the method comprising: sorting the mammalian uterine smooth muscle cells into Lin (CD31, CD45, and Glycophorin A)-positive cells (Lin+) and Lin-negative cells (Lin-) via flow cytometry, isolating sampling the Lin-negative cells, and
further isolating CD34-positive and CD49f-positive cells from the Lin-negative cells.
[claim5]
5. The method according to claim 1, further comprising isolating CD34-positive and CD49f-positive cells from the mammalian uterine smooth muscle cells via flow cytometry.
[claim6]
6. A method for isolating a population of uterine smooth muscle-derived stem cells from mammalian uterine smooth muscle, the method comprising: sampling CD45-negative cells from mammalian uterine smooth muscle via flow cytometry; and
further isolating CD34-positive and CD49f-positive cells from the CD45-negative cells.
[claim7]
7. The method according to claim 1, wherein the mammalian uterine smooth muscle cells are human smooth muscle cells.
[claim8]
8. An isolated population of uterine smooth muscle-derived stem cells, which are CD31-, CD45-, and Glycophorin A-negative and CD34- and CD49f-positive and which are isolated by a method comprising sorting mammalian uterine smooth muscle cells obtained from mammalian uterine smooth muscle into Lin (CD31, CD45, and Glycophorin A)-positive cells (Lin+) and Lin-negative cells (Lin-) via flow cytometry, sampling the Lin-negative cells, and
further isolating CD34-positive and CD49f-positive cells from the Lin-negative cells to obtain the population of uterine smooth muscle-derived stem cells that bind to the anti-CD-34 antibody and to the anti-CD49f antibody.
[claim9]
9. An isolated population of uterine smooth muscle-derived stem cells, which are CD34- and CD49f-positive and which are isolated by a method comprising bringing mammalian uterine smooth muscle cells dispersed from smooth muscle into contact with an anti-CD34 antibody and an anti-CD49f antibody labeled with fluorescent dyes; and
isolating a population of uterine smooth muscle-derived stem cells that bind to the anti-CD34 antibody and the anti-CD49f antibody.
[claim10]
10. An isolated population of uterine smooth muscle-derived stem cells, which are CD45-negative and CD34- and CD49f-positive and which are isolated by a method comprising bringing mammalian uterine smooth muscle cells obtained from mammalian uterine smooth muscle into contact with an anti-CD45 antibody, an anti-CD34 antibody, and an anti-CD49f antibody labeled with fluorescent dyes; and
isolating a population of uterine smooth muscle-derived stem cells that do not bind to the anti-CD45 antibody but bind to the anti-CD34 antibody and the anti-CD49f antibody.
[claim11]
11. The isolated population of uterine smooth muscle-derived stem cells according to claim 9, which are further ABCG2-positive.
[claim12]
12. The isolated population of uterine smooth muscle-derived stem cells according to claim 10, which differentiates into smooth muscle upon transplantation thereof to smooth muscle.
[claim13]
13. The isolated population of uterine smooth muscle-derived stem cells according to claim 12 derived from uterine muscle, which differentiates into uterine muscle upon transplantation thereof into uterine muscle.
[claim14]
14. A composition, comprising the isolated population of uterine smooth muscle-derived stem cells according to claim 9, suitable for regenerating smooth muscle tissue.
[claim15]
15. A method for isolating a population of uterine smooth muscle-derived stem cells from mammalian uterine smooth muscle, the method comprising: bringing the mammalian uterine smooth muscle into contact with an anti-CD31 antibody, an anti-CD45 antibody, an anti-Glycophorin A antibody, an anti-CD34 antibody, and an anti-CD49f antibody labeled with fluorescent dyes; and
isolating a population of uterine smooth muscle-derived stem cells that do not bind to the anti-CD31 antibody, the anti-CD45 antibody, and the anti-Glycophorin A antibody but bind to the anti-CD34 antibody and the anti-CD49f antibody.
[claim16]
16. A method for inducing differentiation of an isolated population of uterine smooth muscle-derived stem cell into fat cells, bone cells, or cartilage cells, the method comprising bringing mammalian uterine smooth muscle cells dispersed from smooth muscle into contact with an anti-CD34 antibody and an anti-CD49f antibody that are labeled with fluorescent dyes,
isolating a population of uterine smooth muscle-derived stem cells that bind to the anti-CD34 antibody and the anti-CD49f antibody, and
culturing the isolated population of uterine smooth muscle-derived stem cells in a medium suitable for inducing differentiation into fat cells, bone cells, or cartilage cells.
[claim17]
17. The method according to claim 16, wherein the isolated smooth muscle-derived stem cells are cultured in a fat cell differentiation medium, osteoblast differentiation medium or cartilage cell differentiation medium.
  • 発明者/出願人(英語)
  • MARUYAMA TETSUO
  • ONO MASANORI
  • KEIO UNIVERSITY
国際特許分類(IPC)
米国特許分類/主・副
  • 424/93.7
  • 435/7.21
  • 435/325
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