Patent SG-11201908905S-A (SG)
Title: Cryopreservation method
Inventor: REID LOLA M
IPC: A01N1/02
Filed: 2018-04-04
Abstract: B00 9140 80,00 ._= =-• 7000 50,00 M1A0 30,00 20,00 10 ,00 00 SOL 1 SOL IA SOL 3 NO CPO MEDIUM 0 10.00 9.00 8.00 1.00 6.00 .7:: 5.00 M 4.00 3.00 2.00 1.00 0.00 SOLI SOL 3 NO CRY0 MEDIUM A 10, 0, 00 0 9 —80,0 60,0 50,00 4000 • = 1 30,00 20,00 10,00 0,0 SOLI 50L3 NO MO MEDIUM E 45 E 40 35 =s 330 25 nl.:\" 20 5 CTRL SOL MO SOL 2A SOL 2B 01.3 1 NO MEDIUM C 1.20 1.00 5.0.80 MI.60 9 0.40 :F2 0.20 0.00 (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (43) International Publication Date 11 October 2018 (11.10.2018) WIPO I PCT Iiiimmomitiaotiolomolmomomil moiflom (10) International Publication Number WO 2018/187439 Al (51) International Patent Classification: A01N 1/02 (2006.01) A61K 47/20 (2006.01) A61K 38/38 (2006.01) C12N 5/00 (2006.01) (21) International Application Number: PCT/US2018/026038 (22) International Filing Date: 04 April 2018 (04.04.2018) (25) Filing Language: English (26) Publication Language: English (30) Priority Data: 62/482,644 06 April 2017 (06.04.2017) US (71) Applicants: THE UNIVERSITY OF NORTH CAR- OLINA AT CHAPEL HILL [US/US]; Office of Com- mercialization and Economic Development, 109 Church Street, Chapel Hill, North Carolina 27516 (US). SAPIEN- ZA UNIVERSITA DI ROMA [IT/IT]; Piazzale Aldo Mo- ro, No. 9, 00185 Roma (IT). (72) Inventors: REID, Lola M.; 3621 Sweeten Creek Road, Chapel Hill, North Carolina 27514 (US). DOMENICO, Al- varo; Via Giuseppe Vasi 18, 00162 Rome (IT). CARDI- NALE, Vincenzo; Viale Regina Margherita, 00171 Rome (IT). GAUDIO, Eugenio; Via G. Tuccimei, 1, 00197 Rome (IT). CARPINO, Guido; Viale Arrigo Boito, 61, 00199 Rome (IT). (74) Agent: VILLACORTA, Gilberto M. et al.; FOLEY & LARDNER LLP, 3000 K Street NM., Suite 600, Washing- ton, District of Columbia 20007-5109 (US). (81) Designated States (unless otherwise indicated, for every kind of national protection available): AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, (54) Title: CRYOPRESERVATION METHOD 1-1 N co 00 O C SOL 1 SOL 3 MEDIUM FIGS. 1A-1E (57) : Human biliary tree stem/progenitors (hBTSCs) are being used for cell therapies of patients with liver cirrhosis. A cryopreservation method was established to optimize sourcing of hBTSCs for these clinical programs and that comprises serum-free Kubota's Medium (KM) supplemented with 10% dimethyl sulfoxide (DMSO), -3% recombinant human albumin and 0.1% hyaluronans. Cryopreserved versus freshly isolated hBTSCs were similar in vitro with respect to self-replication, sternness traits, and multipotency. They were able to differentiate to functional hepatocytes, cholangiocytes or pancreatic islets, yielding similar levels of secretion of albumin or of glucose-inducible levels of insulin. Cryopreserved versus freshly isolated hBTSCs were equally able to engraft into immunocompromised mice yielding cells with human-specific gene expression and human albumin levels in murine serum that were higher for cryopreserved than for freshly isolated hBTSCs. The successful cryoypreservation of hBTSCs facilitates establishment of [Continued on next page] WO 2018/187439 Al IMEDIMOMOIDEIREI0131MOBIEHOMEMOIMIE CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, 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, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. (84) Designated States (unless otherwise indicated, for every 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, TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG). Declarations under Rule 4.17: as to applicant's entitlement to apply for and be granted a patent (Rule 4.17(11)) Published: with international search report (Art. 21(3)) before the expiration of the time limit for amending the claims and to be republished in the event of receipt of amendments (Rule 48.2(h)) hBTSCs cell banking offering logistical advantages for clinical programs for treatment of liver disease.