Generation, expansion and functional analysis of endothelial cells and pericytes derived from human pluripotent stem cells

DOI: 10.1038/nprot.2014.102

Authors: Valeria V Orlova; Francijna E van den Hil; Sandra Petrus-Reurer; Yvette Drabsch; Peter ten Dijke; Christine L Mummery

Step 1: iPSC/ESC maintainance

day 0 ~ 3

culture “adherent” human PSC in “mTeSR” for “3” 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 “3” days.

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

Step 2: mesoderm 1

day 3 ~ 5

culture “adherent” human PSC in “BPEL” for “2” 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 “2” days.

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

Step 3: endothelial 1

day 5 ~ 9

culture “adherent” human PSC in “BPEL” for “4” 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 “4” days.

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

Step 4: endothelial 2

day 9 ~ 11

culture “adherent” human PSC in “BPEL” for “2” 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 “2” days.

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