Patent SG-11201807487W-A (SG)
Title: Generation of midbrain-specific organoids from human pluripotent stem cells
Inventor: JO JUNGHYUN
IPC: C12N5/0793
Filed: 2017-03-14
Abstract: INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property -, Organization 111111110111101110101011111010111110111011110011010111111111111101111011111 International Bureau 0.. .1 j ..... ..or::,„, (10) International Publication Number (43) International Publication Date WO 2017/160234 Al 21 September 2017 (21.09.2017) WIP0 I PCT _ (51) (21) (22) (25) Filing Language: (26) Publication Language: (30) (71) (72) Inventors: JO, Junghyun; c/o Genome Institute of Singa- International Patent Classification: (74) Agent: SPRUSON & FERGUSON (ASIA) PTE LTD; C12N 5/0793 (2010.01) P.O. Box 1531, Robinson Road Post Office, Singapore International Application Number: 903031 (SG). PCT/SG2017/050126 (81) Designated States (unless otherwise indicated, for every kind of national protection available): AE, AG, AL, AM, International Filing Date: AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, 14 March 2017 (14.03.2017) BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, English DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JP, KE, KG, KH, KN, English KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, Priority Data: MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, 10201601965P 14 March 2016 (14.03.2016) SG NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, Applicants: AGENCY FOR SCIENCE, TECHNO- TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, LOGY AND RESEARCH [SG/SG]; 1 Fusionopolis Way, ZA, ZM, ZW. #20-10 Connexis North Tower, Singapore 138632 (SG). NATIONAL UNIVERSITY OF SINGAPORE [SG/SG]; (84) Designated States (unless otherwise indicated, for every 21 Lower Kent Ridge Road, Singapore 119077 (SG). kind of regional protection available): ARIPO (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, pore, 60 Biopolis Street, #02-01, Genome, Singapore TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, 138672 (SG). NG, Huck Hui; c/o Genome Institute of DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, Singapore, 60 Biopolis Street, #02-01, Genome, Singapore LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, 138672 (SG). JE, Hyunsoo Shawn; c/o Duke-NUS Med- SM, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, ical School, 8 College Road, Level 5, #05-17, Singapore GW, KM, ML, MR, NE, SN, TD, TG). 169857 (SG). Declarations under Rule 4.17: — of inventorship (Rule 4.1 7(iv)) [Continued on next page] — _ = — — = (54) Title: GENERATION OF MIDBRAIN-SPECIFIC ORGANOIDS FROM HUMAN PLURIPOTENT STEM CELLS FIG. I A = = = = r Poor piau , mf9urt•on a Organo.d ce,-.1opment = ' , a:L. il V dr...I. '':b. , .1v :.,:.,. Velt\" : , l,t Ir.:1..c: ',Awl - 9 i,•: , ,k. ,- . • .,, - = = unitormel t: ., :1 0 000 c.d1 , . = = = _ : The present disclosure provides a (a) culturing pluripotent stem cells to obtain neuronal from (a) to obtain midbrain regionalized tissues; (c) embedding and culturing matrix to obtain neuroepithelial tissues; and (c) culturing the neuroepithelial Also disclosed herein are culture media suitable for deriving and maintaining neuronal TGF-13 Inhibitor and/or SMAD2/3 inhibitors; and midbrain regionalized tissues comprising (a) signaling protein; and (d) fibroblast growth comprising (a) hedgehog signaling protein; and (b) fibroblast growth a midbrain-like organoid comprising (a) neurotrophin tracellular ing N hedgehog taming ', e.c7ociern , method of deriving and maintaining lineage embryoid (b) WNT-signaling activator; culture media suitable for deriving and maintain - TGF-13 Inhibitor and/or factor; culture media factor; (b) ascorbic ri r ,, a bodies; (b) culturing the neuronal lineage embryoid bod- the midbrain SMAD2/3 inhibitors; suitable for deriving factor and culture media suitable for deriving and main - acid; tissues from (c) to obtain a midbrain-like or - \"I c, midbrain-like organoid in culture, compris - regionalized tissues from (b) in an ex - lineage embryoid bodies comprising (b) WNT-signaling activator; (c) and maintaining neuroepithelial tis - and (c) activator of cAMP-dependent path - 11 7r en (57) N ing e ies I-1 ---.. ganoid. ' L ---- (a) 0 sues .,- way. WO 2017/160234 Al 1#1101MOHIMIRME DEM1100110111100IMMEHOIS • \"\"\"°\"\"`' — with sequence listing part of description (Rule 5.2(a)) — with international search report (Art. 21(3))