Method for inducing vascular endothelial cells
Provided is a method for producing vascular endothelial cells from pluripotent stem cells, the method comprising the following steps (i) to (iii): (i) a step of culturing pluripotent stem cells in a culture medium comprising a BMP, on a culture vessel coated with a first matrix, to produce mesodermal progenitor cells; (ii) a step of dissociating the resulting cells into single cells; and (iii) a step of culturing the resulting cells in a culture medium comprising VEGF, on a culture vessel coated with a second matrix selected from the group consisting of laminin-411 or a fragment thereof, laminin-511 or a fragment thereof, Matrigel, type IV collagen and fibronectin.
1. A method for producing vascular endothelial cells from pluripotent stem cells, the method comprising:
(i) culturing pluripotent stem cells in a culture medium comprising a BMP, on a culture vessel coated with a first matrix, to produce mesodermal progenitor cells;
(ii) dissociating the mesodermal progenitor cells obtained in the culturing of the pluripotent stem cells into single cells; and
(iii) culturing the cells obtained by the dissociating of the mesodermal progenitor cells in a culture medium comprising VEGF, on a culture vessel coated with a second matrix selected from the group consisting of laminin-411 or a fragment thereof having an avidity for integrin, and, laminin-511 or a fragment thereof having an avidity for integrin.
2. The method according to claim 1 , wherein the second matrix used in the culturing of the cells obtained by the dissociating of the mesodermal progenitor cells is a fragment of laminin-411 having an avidity for integrin.
3. The method according to claim 1 , wherein the fragment of laminin-411 is laminin-411 E8.
4. The method according to claim 1 , wherein the first matrix used in the culturing of the pluripotent stem cells is Matrigel, or laminin-511 or a fragment thereof having an avidity for integrin.
5. The method according to claim 4 , wherein the fragment of laminin-511 used in culturing of the pluripotent stem cells is laminin-511 E8.
6. The method according to claim 1 , wherein the BMP is BMP4.
7. The method according to claim 1 , wherein the culture medium used in the culturing of the pluripotent stem cells further comprises a GSK3β inhibitor and VEGF.
8. The method according to claim 7 , wherein the GSK3β inhibitor is CHIR99021.
9. The method according to claim 1 , wherein the culturing of the pluripotent stem cells is carried out for two days or three days.
10. The method according to claim 1 , wherein the culturing of the cells obtained by the dissociating of the mesodermal progenitor cells is carried out for four days.