Functional characterization of iPSC-derived arterial- and venous-like endothelial cells

DOI: 10.1038/s41598-019-40417-9

Authors: S. Rosa; C. Praça; P. R. Pitrez; P. José Gouveia; X. L. Aranguren; L. Ricotti; L. Silva Ferreira

Step 1: iPSC/ESC maintainance

day 0 ~ 1 (duration missing in database; assumed 1 day)

culture “adherent” human PSC in “CDM”. For ECM, use “FN1” at “N/A” “N/A”. Medium should contain factors shown in the following table.

Step 2: mesoderm 1

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

culture “adherent” human PSC in “CDM”. For ECM, use “FN1” at “N/A” “N/A”. Medium should contain factors shown in the following table.

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

Step 3: mesoderm 2

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

culture “adherent” human PSC in “CDM”. For ECM, use “FN1” at “N/A” “N/A”. Medium should contain factors shown in the following table.

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

Step 4: endothelial 1

day 3 ~ 8

culture “adherent” human mesoderm in “CDM” for “5” days. For ECM, use “FN1” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “5” days.

Ideally, “EPC” should express “PECAM1”, “KDR”, “CDH5”, “EphB4” at the end of culture.

Step 5: endothelial 2

day 8 ~ 18

culture “adherent” human EPC in “ESFM” for “10” days. For ECM, use “Gelatin” at “0.1” “%”. Medium should contain factors shown in the following table, and should be changed once in “10” days.

Ideally, “EC” should express “PECAM1”, “KDR”, “CDH5”, “VWF”, “EphB2”, “EFNB2”, “NOTCH4”, “JAG1”, “HEY2”, “DLL4”, “CXCR4”, “EFNB1” at the end of culture.