Establishment of a translational endothelial cell model using directed differentiation of induced pluripotent stem cells from Cynomolgus monkey

DOI: 10.1038/srep35830

Authors: Eva C. Thoma; Tobias Heckel; David Keller; Nicolas Giroud; Brian Leonard; Klaus Christensen; Adrian Roth; Cristina Bertinetti-Lapatki; Martin Graf; Christoph Patsch

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” Cynomolgus monkey PSC in “MT medium” 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 ~ 4

culture “adherent” Cynomolgus monkey PSC in “N2B27 medium” for “3” 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 “3” days.

Ideally, “mesoderm” should express “TBXT” at the end of culture.

Step 3: endothelial 1

day 4 ~ 6

culture “adherent” Cynomolgus monkey mesoderm in “StemPro34-SFM” for “2” 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 “2” days.

Ideally, “EC” should express “CDH5”, “PECAM1”, “VWF”, “KDR” at the end of culture. Ideally, “Pericyte/SMC” should express “PDGFRB” at the end of culture.

Step 4: endothelial 2

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

culture “adherent” Cynomolgus monkey EC in “StemPro34-SFM”. For ECM, use “FN1” at “N/A” “N/A”. Medium should contain factors shown in the following table.