Title:
METHOD FOR PRODUCING PLURIPOTENT STEM CELL-DERIVED INTESTINAL ORGANOID
Document Type and Number:
WIPO Patent Application WO/2020/091020
Kind Code:
A1
Abstract:
The present invention addresses the problem of producing an intestinal organoid, having characteristics more similar to a small intestine of a living body, from pluripotent stem cells. The intestinal organoid is produced from pluripotent stem cells by: (1) a step of differentiating the pluripotent stem cells into endoderm-like cells; (2) a step of differentiating the endoderm-like cells obtained in the step (1) into intestinal stem cell-like cells; (3) a step of culturing the intestinal stem cell-like cells obtained in the step (2) in the presence of an epidermal growth factor, a fibroblast growth factor, a TGFβ receptor inhibitor, a GSK-3β inhibitor and a ROCK inhibitor; (4) a step of forming a spheroid by culturing the cells after the step (3); (5) a step of forming the intestinal organoid by differentiating the spheroid formed in the step (4), wherein the intestinal organoid is produced by a step including a cultivation in the presence of an epidermal growth factor, a BMP inhibitor and a Wnt signal activator. Additionally, the cells constituting the intestinal organoid formed in the step (5) are subjected to planar culture in the presence of an epidermal growth factor and a TGFβ receptor inhibitor to produce a planar culture system. Gas-liquid culture is used during the planar culture to build a high-function evaluation system having a villi structure.
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Inventors:
MATSUNAGA TAMIHIDE (JP)
IWAO TAKAHIRO (JP)
ONOZATO DAICHI (JP)
OGAWA ISAMU (JP)
IWAO TAKAHIRO (JP)
ONOZATO DAICHI (JP)
OGAWA ISAMU (JP)
Application Number:
PCT/JP2019/042938
Publication Date:
May 07, 2020
Filing Date:
October 31, 2019
Export Citation:
Assignee:
UNIV NAGOYA CITY PUBLIC UNIV CORP (JP)
International Classes:
C12N5/071; A61L27/38; C12N5/0735; C12Q1/02
Domestic Patent References:
| WO2017205511A1 | 2017-11-30 | |||
| WO2018038042A1 | 2018-03-01 | |||
| WO2018207714A1 | 2018-11-15 | |||
| WO2014132933A1 | 2014-09-04 | |||
| WO2012060315A1 | 2012-05-10 | |||
| WO2016093222A1 | 2016-06-16 |
Foreign References:
| JP2018207108A | 2018-12-27 | |||
| JP2019092336A | 2019-06-13 | |||
| JP2006239169A | 2006-09-14 |
Other References:
ONOZATO D. ET AL.: "Generation of intestinal organoids suitable for pharmacokinetic studies from human induced pluripotent stem cells.", DRUG METAB DISPOS., vol. 46, no. 11, 3 April 2018 (2018-04-03), pages 1572 - 1580, XP055659658, DOI: 10.1124/dmd.118.080374
ONOZATO D. ET AL.: "Efficient generation of cynomologous monkey induced pluripotent stem cell -derived intestinal organoids with pharmacokinetic functions", STEM CELLS AND DEVELOPMENT, vol. 27, no. 15, 10 May 2018 (2018-05-10), pages 1033 - 1045, XP055659656
ZHANG R. ET AL.: "Human iPSC-derived posterior gut progenitors are expandable and capable of forming gut and liver organoids", STEM CELL REPORTS, vol. 10, 13 March 2018 (2018-03-13), pages 780 - 793, XP055690711, DOI: 10.1016/j.stemcr.2018.01.006
KABEYA T. ET AL.: "Cyclic AMP signaling promotes the differentiation of human induced pluripotent stem cells into intestinal epithelial cells", DRUG MATAB DISPOS, vol. 46, no. 10, 1 August 2018 (2018-08-01), pages 1411 - 1419, XP055629232, DOI: 10.1124/dmd.118.082123
DRUG METAB. PHARMACOKINETIC, vol. 29, no. 1, 2014, pages 44 - 51
NATURE, vol. 470, 2011, pages 105 - 109
ZHANG X. ET AL., STEM CELLS, vol. 24, 2006, pages 2669 - 2676
BIOCHEM. BIOPHYS. RES. COMMUN., vol. 391, no. 7, 2010, pages 38 - 42
"Manipulating the Mouse Embryo A Laboratory Manual", 1994, COLD SPRING HARBOR LABORATORY PRESS
CIBELLI ET AL., SCIENCE, vol. 280, 1998, pages 1256 - 1147
WILMUT ET AL., NATURE, vol. 385, 1997, pages 810
AKIRA IRITANI ET AL., PROTEIN NUCLEIC ACID ENZYME, vol. 44, 1999, pages 892
BAGUISI ET AL., NATURE BIOTECHNOLOGY, vol. 17, 1999, pages 456
WAKAYAMA ET AL., NATURE, vol. 394, 1998, pages 369
NATURE GENETICS, vol. 22, 1999, pages 127
PROC. NATL. ACAD. SCI. USA, vol. 96, 1999, pages 14984
RIDEOUT III ET AL., NATURE GENETICS, vol. 24, 2000, pages 109
TACHIBANA ET AL.: "Human Embryonic Stem Cells Derived by Somatic Cell Nuclear Transfer", CELL, vol. 153, 2013, pages 1228 - 1238, XP055295481, DOI: 10.1016/j.cell.2013.05.006
YU J ET AL., SCIENCE, vol. 318, no. 5858, 2007, pages 1917 - 1920
NAKAJIMA ET AL., STEM CELLS, vol. 25, 2007, pages 983 - 985
KIM ET AL., CELL STEM CELL, vol. 1, 2007, pages 346 - 352
REVAZOVA, CLONING STEM CELLS, vol. 9, 2007, pages 432 - 449
REVAZOVA ET AL., CLONING STEM CELLS, vol. 10, 2008, pages 11 - 24
STRELCHENKO N. ET AL., REPROD BIOMED ONLINE, vol. 9, 2004, pages 623 - 629
KLIMANSKAYA I. ET AL., NATURE, vol. 444, 2006, pages 481 - 485
CHUNG Y. ET AL., CELL STEM CELL, vol. 2, 2008, pages 113 - 117
WASSARMAN, P.M. ET AL., METHODS IN ENZYMOLOGY, vol. 365, 2003
MATSUI ET AL., CELL, vol. 70, 1992, pages 841 - 847
SHAMBLOTT ET AL., PROC. NATL. ACAD. SCI. USA, vol. 95, no. 23, 1998, pages 13726 - 13731
TURNPENNY ET AL., STEM CELLS, vol. 21, no. 5, 2003, pages 598 - 609
TAKAHASHI KYAMANAKA S, CELL, vol. 126, no. 4, 2006, pages 663 - 676
TAKAHASHI K ET AL., CELL, vol. 131, no. 5, 2007, pages 861 - 72
NAKAGAWA M ET AL., NAT. BIOTECHNOL., vol. 26, no. 11, 2008, pages 1269 - 1275
KIM JB ET AL., NATURE, vol. 454, no. 7204, 2008, pages 646 - 650
KIM JB ET AL., CELL, vol. 136, no. 3, 2009, pages 411 - 419
KIM DKIM CHMOON JI ET AL., CELL STEM CELL, vol. 4, 2009, pages 472 - 476
SILVA J ET AL., PLOS. BIOL., vol. 6, no. 10, 2008, pages e253
STADTFELD M ET AL., SCIENCE, vol. 322, no. 5903, 2008, pages 949 - 953
WOLTJEN KMICHAEL IPMOHSENI P ET AL., NATURE, vol. 458, 2009, pages 771 - 775
YUSA KRAD RTAKEDA J ET AL., NAT METHODS, vol. 6, 2009, pages 363 - 369
YU JHU KSMUGA-OTTO KTIAN S ET AL., SCIENCE, vol. 324, 2009, pages 797 - 801
CAS, no. 1135306-29-4
ONOZATO D. ET AL.: "Efficient generation of cynomologous monkey induced pluripotent stem cell -derived intestinal organoids with pharmacokinetic functions", STEM CELLS AND DEVELOPMENT, vol. 27, no. 15, 10 May 2018 (2018-05-10), pages 1033 - 1045, XP055659656
ZHANG R. ET AL.: "Human iPSC-derived posterior gut progenitors are expandable and capable of forming gut and liver organoids", STEM CELL REPORTS, vol. 10, 13 March 2018 (2018-03-13), pages 780 - 793, XP055690711, DOI: 10.1016/j.stemcr.2018.01.006
KABEYA T. ET AL.: "Cyclic AMP signaling promotes the differentiation of human induced pluripotent stem cells into intestinal epithelial cells", DRUG MATAB DISPOS, vol. 46, no. 10, 1 August 2018 (2018-08-01), pages 1411 - 1419, XP055629232, DOI: 10.1124/dmd.118.082123
DRUG METAB. PHARMACOKINETIC, vol. 29, no. 1, 2014, pages 44 - 51
NATURE, vol. 470, 2011, pages 105 - 109
ZHANG X. ET AL., STEM CELLS, vol. 24, 2006, pages 2669 - 2676
BIOCHEM. BIOPHYS. RES. COMMUN., vol. 391, no. 7, 2010, pages 38 - 42
"Manipulating the Mouse Embryo A Laboratory Manual", 1994, COLD SPRING HARBOR LABORATORY PRESS
CIBELLI ET AL., SCIENCE, vol. 280, 1998, pages 1256 - 1147
WILMUT ET AL., NATURE, vol. 385, 1997, pages 810
AKIRA IRITANI ET AL., PROTEIN NUCLEIC ACID ENZYME, vol. 44, 1999, pages 892
BAGUISI ET AL., NATURE BIOTECHNOLOGY, vol. 17, 1999, pages 456
WAKAYAMA ET AL., NATURE, vol. 394, 1998, pages 369
NATURE GENETICS, vol. 22, 1999, pages 127
PROC. NATL. ACAD. SCI. USA, vol. 96, 1999, pages 14984
RIDEOUT III ET AL., NATURE GENETICS, vol. 24, 2000, pages 109
TACHIBANA ET AL.: "Human Embryonic Stem Cells Derived by Somatic Cell Nuclear Transfer", CELL, vol. 153, 2013, pages 1228 - 1238, XP055295481, DOI: 10.1016/j.cell.2013.05.006
YU J ET AL., SCIENCE, vol. 318, no. 5858, 2007, pages 1917 - 1920
NAKAJIMA ET AL., STEM CELLS, vol. 25, 2007, pages 983 - 985
KIM ET AL., CELL STEM CELL, vol. 1, 2007, pages 346 - 352
REVAZOVA, CLONING STEM CELLS, vol. 9, 2007, pages 432 - 449
REVAZOVA ET AL., CLONING STEM CELLS, vol. 10, 2008, pages 11 - 24
STRELCHENKO N. ET AL., REPROD BIOMED ONLINE, vol. 9, 2004, pages 623 - 629
KLIMANSKAYA I. ET AL., NATURE, vol. 444, 2006, pages 481 - 485
CHUNG Y. ET AL., CELL STEM CELL, vol. 2, 2008, pages 113 - 117
WASSARMAN, P.M. ET AL., METHODS IN ENZYMOLOGY, vol. 365, 2003
MATSUI ET AL., CELL, vol. 70, 1992, pages 841 - 847
SHAMBLOTT ET AL., PROC. NATL. ACAD. SCI. USA, vol. 95, no. 23, 1998, pages 13726 - 13731
TURNPENNY ET AL., STEM CELLS, vol. 21, no. 5, 2003, pages 598 - 609
TAKAHASHI KYAMANAKA S, CELL, vol. 126, no. 4, 2006, pages 663 - 676
TAKAHASHI K ET AL., CELL, vol. 131, no. 5, 2007, pages 861 - 72
NAKAGAWA M ET AL., NAT. BIOTECHNOL., vol. 26, no. 11, 2008, pages 1269 - 1275
KIM JB ET AL., NATURE, vol. 454, no. 7204, 2008, pages 646 - 650
KIM JB ET AL., CELL, vol. 136, no. 3, 2009, pages 411 - 419
KIM DKIM CHMOON JI ET AL., CELL STEM CELL, vol. 4, 2009, pages 472 - 476
SILVA J ET AL., PLOS. BIOL., vol. 6, no. 10, 2008, pages e253
STADTFELD M ET AL., SCIENCE, vol. 322, no. 5903, 2008, pages 949 - 953
WOLTJEN KMICHAEL IPMOHSENI P ET AL., NATURE, vol. 458, 2009, pages 771 - 775
YUSA KRAD RTAKEDA J ET AL., NAT METHODS, vol. 6, 2009, pages 363 - 369
YU JHU KSMUGA-OTTO KTIAN S ET AL., SCIENCE, vol. 324, 2009, pages 797 - 801
CAS, no. 1135306-29-4
Attorney, Agent or Firm:
HAGINO Mikiharu (JP)
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