Method for producing intestinal cells
An object of the present invention is to provide a method of producing intestinal cells by use of pluripotent stem cells as a starting material. According to the present invention, provided is a method of producing intestinal cells, comprising the steps of: (A) inducing differentiation of pluripotent stem cells into definitive endoderm cells; and (B) culturing the definitive endoderm cells in the presence of (2′Z,3′E)-6-bromoindirubin-3′-oxime (BIO) and N-[(3,5-difluorophenyl)acetyl]-L-Ala-2-phenyl-L-Gly-tert-butyl-OH (DAPT) to thereby induce differentiation of the definitive endoderm cells into intestinal cells.
1. A method of producing intestinal cells expressing Cdx2, comprising the steps of:
(a) obtaining mouse or human embryonic stem (ES) cells,
(b) culturing the ES cells of step (a) with Activin and/or bFGF to produce definitive endoderm cells,
(c) culturing the definitive endoderm cells of step (b) with (2′Z,3′E)-6-bromoindirubin-3′-oxime (BIO) and N-[(3,5-difluorophenyl)acetyl]-L-Ala-2-phenyl-L-Gly-tert-butyl-OH (DAPT) and in the presence of M15 cells or MEF cells, to thereby induce differentiation of the definitive endoderm cells into intestinal cells expressing Cdx2.
2. The method according to claim 1 , wherein the definitive endoderm cells are separated from a cell culture obtained in step (b) by flow cytometry using fluorescently-labelled antibodies against E-cadherin (ECD) and CXCR4, and said separated definitive endoderm cells are used in the step (c).
3. The method according to claim 1 , wherein the ES cells are cultured in the presence of feeder cells.
4. The method according to claim 3 , wherein the feeder cells are cells derived from a mesoderm.