Differentiation and characterization of human iPSC-derived vascular endothelial cells under physiological shear stress

DOI: 10.1016/j.xpro.2021.100394

Authors: Nadia O. Abutaleb; George A. Truskey

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “mTeSR Plus” for “1” days. For ECM, use “Matrigel” at “1” “%”. 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: Primitive streak 1

day 1 ~ 2 (duration missing in database; assumed 1 day)

culture “adherent” human PSC in “unspecified medium”. For ECM, use “Matrigel” at “1” “%”.

Step 3: Primitive streak 2

day 2 ~ 3 (duration missing in database; assumed 1 day)

culture “adherent” unknown species cell in “unspecified medium”. For ECM, use “Matrigel” at “1” “%”.

Step 4: mesoderm 1

day 3 ~ 6

culture “adherent” human PSC in “N2B27 medium” for “3” days. For ECM, use “Matrigel” at “1” “%”. Medium should contain factors shown in the following table, and should be changed once in “3” days.

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

Step 5: mesoderm 2

day 6 ~ 7 (duration missing in database; assumed 1 day)

culture “adherent” unknown species cell in “unspecified medium”. For ECM, use “Matrigel” at “1” “%”.

Step 6: endothelial 1

day 7 ~ 8

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

Step 7: endothelial 2

day 8 ~ 10

culture “adherent” unknown species cell in “StemPro” for “2” days. For ECM, use “Collagen” at “0.05” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

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