The influence of a spatiotemporal 3D environment on endothelial cell differentiation of human induced pluripotent stem cells

DOI: 10.1016/j.biomaterials.2014.01.037

Authors: Sophia Zhang; James R. Dutton; Liping Su; Jianyi Zhang; Lei Ye

Step 1: iPSC/ESC maintainance

day 0 ~ 2

culture “spheroid” human PSC in “unspecified medium” for “2” days. “PSC” should be “upregulated” for “GFP”. For ECM, use “fibrin” at “N/A” “N/A”. Medium should be changed once in “2” days.

Step 2: mesoderm 1

day 2 ~ 3

culture “spheroid” 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, “mesoderm” should be obtained at the end of culture.

Step 3: endothelial 1

day 3 ~ 5

culture “spheroid” 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 4: endothelial 2

day 5 ~ 7

culture “adherent” 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”, “vWF-8” at the end of culture.