The prostaglandin H2 analog U-46619 improves the differentiation efficiency of human induced pluripotent stem cells into endothelial cells by activating both p38MAPK and ERK1/2 signaling pathways

DOI: 10.1186/s13287-018-1061-4

Authors: Liping Su; Xiaocen Kong; Szeyun Lim; Szejie Loo; Shihua Tan; Kiankeong Poh; James Dutton; Colin Stewart; Stuart Cook; Xiaofei Su; Jianhua Ma; Jianyi Zhang; Lei Ye

Step 1: iPSC/ESC maintainance

day 0 ~ 4

culture “spheroid” human PSC in “E8” for “4” days. Medium should contain factors shown in the following table, and should be changed once in “4” days.

Ideally, “PSC” should be obtained at the end of culture.

Step 2: Primitive streak 1

day 4 ~ 5

culture “unspecified method” human PSC in “EBM2” for “1” days. For ECM, use “fibrin” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Step 3: mesoderm 1

day 5 ~ 6

culture “unspecified method” human PSC in “EBM2” for “1” days. For ECM, use “fibrin” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Ideally, “dummy” should express “TBXT”, “Etv2”, “Gata-2”, “Tal-1” at the end of culture.

Step 4: endothelial 1

day 6 ~ 8

culture “unspecified method” human PSC in “EBM2” for “2” days. For ECM, use “fibrin” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Step 5: endothelial 2

day 8 ~ 10

culture “unspecified method” human PSC in “EGM2-MV” for “2” days. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Ideally, “EC” should express “PECAM1”, “CDH5”, “DLL4”, “EphB4”, “CXCR4” at the end of culture.