Differentiation and expansion of endothelial cells requires pre-optimization of KDR+ expression kinetics

DOI: 10.1016/j.scr.2019.101685

Authors: Basharat Jahan; Kara E. McCloskey

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “E8” for “1” days. For ECM, use “Matrigel” at “0.2” “g/m2”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

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

Step 2: mesoderm 1

day 1 ~ 3

culture “adherent” human PSC in “unspecified medium” for “2” days. For ECM, use “FN1” at “0.05” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Ideally, “VPC” should express “KDR” at the end of culture.

Step 3: endothelial 1

day 3 ~ 10

culture “adherent” human PSC in “unspecified medium” for “7” days. For ECM, use “FN1” at “0.05” “g/l”. Medium should be changed once in “7” days.

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

Step 4: endothelial 2

day 10 ~ 13

culture “adherent” human PSC in “unspecified medium” for “3” days. For ECM, use “FN1” at “0.05” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “3” days.

Ideally, “EC” should express “CDH5”, “KDR” at the end of culture.