Endothelial and hematopoietic hPSCs differentiation via a hematoendothelial progenitor

DOI: 10.1186/s13287-022-02925-w

Authors: Alejandra Vargas-Valderrama; Anne-Charlotte Ponsen; Morgane Le Gall; Denis Clay; Sébastien Jacques; Tudor Manoliu; Valérie Rouffiac; Karine Ser-le-Roux; Cyril Quivoron; Fawzia Louache; Georges Uzan; Maria-Teresa Mitjavila-Garcia; Estelle Oberlin; Hind Guenou

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “KnockOut DMEM” for “1” days. 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 ~ 3

culture “spheroid” human PSC in “Stemline II Hematopoietic Stem Cell Expansion Medium” for “2” days. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Step 3: mesoderm 1

day 3 ~ 5

culture “spheroid” human PSC in “Stemline II Hematopoietic Stem Cell Expansion Medium” for “2” days. 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 4: mesoderm 2

day 5 ~ 6

culture “spheroid” human mesoderm in “Stemline II Hematopoietic Stem Cell Expansion Medium” for “1” days. Medium should be changed once in “1” days.

Step 5: endothelial 1

day 6 ~ 8

culture “adherent” human PSC in “STEMdiff APEL2 Medium” for “2” 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 “2” days.

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