Arterial specification of endothelial cells derived from human induced pluripotent stem cells in a biomimetic flow bioreactor

DOI: 10.1016/j.biomaterials.2015.02.121

Authors: Amogh Sivarapatna; Mahboobe Ghaedi; Andrew V. Le; Julio J. Mendez; Yibing Qyang; Laura E. Niklason

Step 1: iPSC/ESC maintainance

day 0 ~ 1

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

Ideally, “PSC” should express “POU5F1”, “SOX2”, “NANOG” at the end of culture.

Step 2: mesoderm 1

day 1 ~ 5

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

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

Step 3: endothelial 1

day 5 ~ 7

culture “adherent” human mesoderm in “VascuLife VEGF medium” for “2” days. For ECM, use “Gelatin” at “0.67” “%”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Ideally, “EC” should express “PECAM1”, “CDH5”, “VWF”, “NOS3”, “CD105”, “KDR” at the end of culture.

Step 4: endothelial 2

day 7 ~ 8

culture “device” human EC in “VascuLife VEGF medium” for “1” days. For ECM, use “FN1” at “0.03” “g/m2”. Medium should be changed once in “1” days.

Ideally, “EC” should express “KDR”, “EphrinB2”, “CXCR4”, “connexin40”, “NOTCH1”, “KLF2”, “KLF4” at the end of culture.