Method of producing a mesodermal-lineage primitive streak cell
A method of producing a mesodermal-lineage primitive streak cell includes a first step of culturing pluripotent stem cells in a medium containing FGF2, BMP (bone morphogenetic protein) 4, a GSK-3β inhibitor, and retinoic acid or a derivative thereof and a second step of culturing cells obtained in the first step in a medium containing FGF2, a GSK-3β inhibitor, and BMP7.
1 . A method of producing a mesodermal-lineage primitive streak cells comprising:
(i) culturing pluripotent stem cells in a medium containing fibroblast growth factor 2 (FGF2), bone morphogenic protein (BMP) 4, a glycogen synthase kinase-3β (GSK-3β) inhibitor, and retinoic acid or a derivative thereof; and
(ii) culturing cells obtained in (i) in a medium containing FGF2, a GSK-3β inhibitor, and BMP7 thereby producing the mesodermal-lineage primitive streak cell.
2 . The method of claim 1 , wherein said mesodermal-lineage primitive streak cell is positive for CDX1 and BRACHYURY.
3 . The method of claim 1 , wherein said GSK-3β inhibitor is CHIR99021.
4 . The method of claim 1 , wherein said culturing in step (i) and said culturing in step (ii) are performed using a culture vessel coated with an extracellular matrix.
5 . The method of claim 4 , wherein the extracellular matrix is laminin 511 E8 fragment.
6 . The method of claim 1 , wherein the pluripotent stem cells are induced pluripotent stem (iPS) cells.
7 . The method of claim 6 , wherein the iPS cells are human iPS cells.