Patent ID: 11859206
Assignee: UNIVERSITY HEALTH NETWORK
Field: Pharmaceuticals (Chemistry)
Classification: CPC A  C | IPC C

Claim 0:
1. A method of producing oligodendrogenic neural progenitor cells (o-NPCs), the method comprising:
a) obtaining iPSCs cultured for at least about 2 days in vessels comprising a gelatinous matrix with an induced pluripotent cell media/embryonic cell media supplemented with a ROCK inhibitor and culturing the iPSCs:
i. in NIM supplemented with leukemia inhibitory factor (LIF), FGF agonist, B27 supplement lacking vitamin A, N2 supplement, TGFb inhibitor, BMP inhibitor for about 7 days; and
ii. in NIM supplemented with EGFR agonist, FGFR agonist, B27 supplement lacking vitamin A and N2 supplement, wherein the iPSCs are cultured in vessels coated with a gelatinous matrix comprising poly-L-lysine/laminin for about 1 to 2 days to produce columnar cells in the form of rosettes expressing Pax 6;

b) culturing the columnar cells in the form of rosettes from step ii. in NEM comprising EGFR agonist, FGFR agonist, B27 lacking vitamin A, and N2 supplement for about 4 days, wherein the columnar cells are cultured in vessels coated with a gelatinous matrix comprising poly-L-lysine/laminin, to produce unpatterned NPCs;
c) culturing the unpatterned NPCs from step b) for about 6 days in NEM comprising retinoic acid, N2 supplement, B27 supplement, EGFR agonist and a Shh agonist to produce caudalized NPCs;
d) culturing the caudalized NPCs from step c):
i. in NEM comprising EGFR agonist, N2 supplement, B27 supplement, retinoic acid and Shh agonist for about 3 to about 6 days; and
ii. in NEM comprising FGFR agonist, N2 supplement, B27 supplement and a Shh agonist for about 3 days to obtain ventralized NPCs;

e) culturing the ventralized NPCs for about 12 to about 16 days in NEM comprising i) PDGFR agonist for the about 12 to about 16 days; ii) B27 and N1 supplements for the preliminary about 12 days; and iii) thyroxine or a thyroxine analogue for the latter about 7 to about 9 days to produce o-NPCs.