| 主权项 |
1. A method for generating a beta cell from a stem cell or an induced pluripotent stem cell, the method comprising:
(a) contacting the cells with a first culture medium, wherein the first culture medium is an RPMI medium comprising 1× Pen-Strep and 1× Glutamax and wherein the first culture medium further comprises Activin A, Wnt3A and Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid, (b) contacting the cells with a second culture medium, wherein the second culture medium is an RPMI medium comprising 1× Pen-Strep and 1× Glutamax and wherein the second culture medium further comprises Activin A protein and FBS in RPMI medium, (c) contacting the cells with a third culture medium, wherein the third culture medium is an RPMI medium comprising 1× Pen-Strep and 1× Glutamax and wherein the third culture medium further comprises containing human FGF10 protein, KAAD-cyclopamine and FBS in RPMI medium, (d) contacting the cells with a fourth culture medium, wherein the fourth culture medium is an DMEM high glucose medium comprising 1× Pen-Strep and 1× Glutamax and wherein the fourth culture medium further comprises FGF 10, KAAD-cyclopamine, retinoic acid, LDN-193189 and 1×B27, (e) contacting the cells with a fifth culture medium, wherein the fifth culture medium is a CMRL medium comprising 1× Pen-Strep and 1× Glutamax and wherein the fourth culture medium further comprises exedin-4, SB431542 and 1×B27, and (f) contacting the cells with a sixth culture medium, wherein the sixth culture medium is a CMRL medium comprising 1× Pen-Strep and 1× Glutamax and wherein the sixth culture medium further comprises 4-(4-Hydroxy-3,5-diiodophenoxy)-3,5-diiodobenzeneacetic acid and 1×B27. |
