(19) TZZ __Z_T (11) EP 2 171 045 B1 (12) EUROPEAN PATENT SPECIFICATION (45) Date of publication and mention (51) Int Cl.: of the grant of the patent: C12N 5/071 (2010.01) C12N 5/074 (2010.01) 21.12.2016 Bulletin 2016/51 (86) International application number: (21) Application number: 08807182.4 PCT/IB2008/002540 (22) Date of filing: 13.06.2008 (87) International publication number: WO 2009/007852 (15.01.2009 Gazette 2009/03) (54) MULTIPOTENT/PLURIPOTENT CELLS AND METHODS MULTIPOTENTE/PLURIPOTENTE ZELLEN UND ENTSPRECHENDE VERFAHREN CELLULES MULTIPOTENTES/PLURIPOTENTES ET PROCÉDÉS S’Y RAPPORTANT (84) Designated Contracting States: WO-A-2009/032194 WO-A-2009/032456 AT BE BG CH CY CZ DE DK EE ES FI FR GB GR WO-A-2009/057831 WO-A-2009/061442 HR HU IE IS IT LI LT LU LV MC MT NL NO PL PT RO SE SI SK TR • JAHAGIRDAR BALKRISHNA N ET AL: "Multipotent adult progenitor cell and stem cell (30) Priority: 15.06.2007 JP 2007159382 plasticity" STEM CELL REVIEWS, HUMANA 20.11.2007 PCT/EP2007/010019 PRESS, US, vol. 1, no. 1, 1 January 2005 28.03.2008 US 40646 (2005-01-01), pages 53-59, XP002478582 ISSN: 1550-8943 (43) Date of publication of application: • D’IPPOLITO G ET AL: "Marrow-isolated adult 07.04.2010 Bulletin 2010/14 multilineage inducible (MIAMI) cells, a unique population of postnatal young and old human (73) Proprietor: Kyoto University cells with extensive expansion and Sakyo-ku differentiation potential" JOURNAL OF CELL Kyoto-shi SCIENCE, CAMBRIDGE UNIVERSITY PRESS, Kyoto 606-8501 (JP) LONDON, GB, vol. 117, no. part 14, 15 June 2004 (2004-06-15), pages 2971-2981, XP002339338 (72) Inventors: ISSN: 0021-9533 • SAKURADA, Kazuhiro • TAKAHASHI K ET AL: "Induction of pluripotent Yokohama 225-0016 (JP) stem cells from mouse embryonic and adult • MASAKI, Hideki fibroblast cultures by defined factors" CELL, Tokyo 108-0071 (JP) CELL PRESS, CAMBRIDGE, NA, US, vol. 126, no. • ISHIKAWA, Tetsuya 4, 25 August 2006 (2006-08-25), pages 663-676, Tokyo 104-0045 (JP) XP003013968 ISSN: 0092-8674 • TAKAHASHI, Shunichi • WERNIGMARIUS ET AL: "In vitro reprogramming Hyogo 657-0064 (JP) of fibroblasts into a pluripotent ES-cell-like state" NATURE, NATURE PUBLISHING GROUP, (74) Representative: Hoffmann Eitle LONDON, UK, vol. 448, no. 7151, 6 June 2007 Patent- und Rechtsanwälte PartmbB (2007-06-06), pages 318-325, XP002468993 ISSN: Arabellastraße 30 0028-0836 81925 München (DE) • OKITA KEISUKE ET AL: "Generation of germline-competent induced pluripotent stem (56) References cited: cells" NATURE, NATURE PUBLISHING GROUP, EP-A- 1 384 775 WO-A-01/21767 LONDON, UK, vol. 448, no. 7151, 6 June 2007 WO-A-2007/069666 WO-A-2008/038148 (2007-06-06), page 313, XP002475653 ISSN: WO-A-2008/118820 WO-A-2008/151058 0028-0836 Note: Within nine months of the publication of the mention of the grant of the European patent in the European Patent Bulletin, any person may give notice to the European Patent Office of opposition to that patent, in accordance with the Implementing Regulations. Notice of opposition shall not be deemed to have been filed until the opposition fee has been paid. (Art. 99(1) European Patent Convention). EP 2 171 045 B1 Printed by Jouve, 75001 PARIS (FR) (Cont. next page) EP 2 171 045 B1 • NIMET MAHERALI ET AL: "Directly • YU JUNYING ET AL: "Induced pluripotent stem Reprogrammed Fibroblasts Show Global cell lines derived from human somatic cells" Epigenetic Remodeling and Widespread Tissue SCIENCE, AMERICAN ASSOCIATION FOR THE Contribution" CELL STEM CELL, CELL PRESS, ADVANCEMENT OF SCIENCE, US, vol. 318, no. US, vol. 1, 1 June 2007 (2007-06-01), pages 55-70, 5858, 21 December 2007 (2007-12-21), pages XP009091847 ISSN: 1934-5909 1917-1920, XP009105055 ISSN: 1095-9203 • HENG B C ET AL: "Incorporating protein • PARK IN-HYUN ET AL: "Reprogramming of transduction domains (PTD) within intracellular human somatic cells to pluripotency with defined proteins associated with the ’stemness’ factors" NATURE, NATURE PUBLISHING phenotype. Novel use of such recombinant GROUP, LONDON, UK, vol. 451, no. 7175, 23 ’fusion’ proteins to overcome current limitations December 2007 (2007-12-23), pages 141-147, of applying autologous adult stem cells in XP002475655 ISSN: 0028-0836 regenerative medicine?" MEDICAL • NAKAGAWA MASATO ET AL: "Generation of HYPOTHESES, EDEN PRESS, PENRITH, US, vol. induced pluripotent stem cells without Myc from 64, no. 5, 1 January 2005 (2005-01-01), pages mouse and human fibroblasts" LASER FOCUS 992-996, XP004788973 ISSN: 0306-9877 WORLD, PENNWELL, TULSA, OK, US, vol. 26, no. • BECKER-HAPAK MICHELLE ET AL: "Protein 1, 1 January 2008 (2008-01-01), pages 101-106, transduction: generation of full-length XP009098334 ISSN: 1043-8092 transducible proteins using the TAT system" • WERNIG MARIUS ET AL: "c-Myc is dispensable CURRENT PROTOCOLS IN CELL BIOLOGY, for direct reprogramming of mouse fibroblasts" JOHN WILEY & SONS, INC, US, vol. 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EXPERT OPINION ON BIOLOGICAL PRESS, CAMBRIDGE, NA, US, vol. 131, no. 5, 30 THERAPY, ASHLEY, LONDON, GB, vol. 6, no. 12, November 2007 (2007-11-30), pages 861-872, 1 December 2006 (2006-12-01), pages 1251-1254, XP002478584 ISSN: 0092-8674 XP009098586 ISSN: 1471-2598 • MASAKI H ET AL: "Heterogeneity of pluripotent • STEWART ET AL: "Mechanisms of self-renewal marker gene expression in colonies generated in in human embryonic stem cells" EUROPEAN human iPS cell induction culture" STEM CELL JOURNAL OF CANCER, PERGAMON PRESS, RESEARCH,, vol. 1, no. 2, 31 January 2008 OXFORD, GB, vol. 42, no. 9, 1 June 2006 (2008-01-31), pages 105-115, XP022764781 ISSN: (2006-06-01), pages 1257-1272, XP005496720 1873-5061 [retrieved on 2008-01-31] ISSN: 0959-8049 • BOSNALI MANAL ET AL: "Generation of • WATANABE KIICHI ET AL: "A ROCK inhibitor transducible versions of transcription factors permits survival of dissociated human Oct4 and Sox2" BIOLOGICAL CHEMISTRY, vol. embryonic stem cells" NATURE 389, no. 7, July 2008 (2008-07), pages 851-861, BIOTECHNOLOGY, NATURE PUBLISHING XP009118395 ISSN: 1431-6730 GROUP, NEW YORK, NY, US, vol. 25, no. 6, 1 June • MAHERALI NIMET ET AL: "Guidelines and 2007 (2007-06-01), pages 681-686, XP002458303 Techniques for the Generation of Induced ISSN: 1087-0156 Pluripotent Stem Cells" CELL STEM CELL, vol. 3, no. 6, December 2008 (2008-12), pages 595-605, XP002532630 ISSN: 1934-5909 2 (Cont. next page) EP 2 171 045 B1 • ZHOU HONGYAN ET AL: "Generation of Induced Pluripotent Stem Cells Using Recombinant Proteins" CELL STEM CELL, vol. 4, no. 5, May 2009 (2009-05), pages 381-384, XP002532629 ISSN: 1934-5909 • HUANGFU DANWEI ET AL: "Induction of pluripotent stem cells from primary human fibroblasts with only Oct4 and Sox2", NATURE BIOTECHNOLOGY, NATURE PUBLISHING GROUP, NEW YORK, NY, US, vol. 26, no. 11, 1 November 2008 (2008-11-01), pages 1269-1275, XP002534948, ISSN: 1087-0156, DOI: 10.1038/NBT.1502 [retrieved on 2008-10-12] 3 EP 2 171 045 B1 Description CROSS-REFERENCE 5 [0001] This application claims the benefit of Japanese Patent Application, JPO 2007 -159382, filed June 15, 2007, PCT/EP2007/010019, filed November 20, 2007, and United States Provisional Application, 61/040,646, filed March 28, 2008. BACKGROUND OF THE INVENTION 10 [0002] The field of regenerative medicine encompasses therapies designed to aid the repair, replacement, or regen- eration of damaged cells, tissues, or organs. One branch of regenerative medicine includes cell therapies that rely on embryonic stem cells (ES), which have the potential to give rise to a diverse range of cell types. ES-based cell therapies have the promise of treating a variety of health conditions including Alzheimer’s Disease, Parkinson’s Disease, stroke, 15 spinal injuries, heart attack, renal failure, osteoporosis, type I diabetes, multiple sclerosis, rheumatoid arthritis, burns, and wounds. However, the progress of such therapies has been hindered by a range of factors including the possibility of immune rejection of ES cells derived from a donor who is immunologically incompatible with the recipient. SUMMARY OF THE INVENTION 20 [0003] The present invention relates to a method as set forth in the claims. [0004] This disclosure encompasses human stem cells that in some cases are pluripotent and in some cases are multipotent. The disclosure further encompasses methods for generating such human stem cells, methods for using such stem cells, and related compositions.