Identification of a retinoic acid-dependent haemogenic endothelial progenitor from human pluripotent stem cells

DOI: 10.1038/s41556-022-00898-9

Authors: Stephanie A. Luff; J. Philip Creamer; Sara Valsoni; Carissa Dege; Rebecca Scarfò; Analisa Dacunto; Sara Cascione; Lauren N. Randolph; Eleonora Cavalca; Ivan Merelli; Samantha A. Morris; Andrea Ditadi; Christopher M. Sturgeon

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “unspecified medium” for “1” days. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should be changed once in “1” days.

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

Step 2: Primitive streak 1

day 1 ~ 2

culture “adherent” human PSC in “unspecified medium” for “1” days. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should be changed once in “1” days.

Step 3: Primitive streak 2

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

culture “spheroid” human PSC in “unspecified medium”. Medium should contain factors shown in the following table.

Step 4: mesoderm 1

day 3 ~ 4 (duration missing in database; assumed 1 day)

culture “spheroid” human PSC in “StemPro-34 medium”. Medium should contain factors shown in the following table.

Ideally, “mesoderm” should express “KDR”, “CXCR4”, “ALDH1A2”, “CYP26A1”, “GYPA”, “CDX1”, “CDX2”, “CDX4”, “TBXT”, “ALDH1A1” at the end of culture.

Step 5: mesoderm 2

day 4 ~ 5

culture “spheroid” human mesoderm in “StemPro-34 medium” for “1” days. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Step 6: endothelial 1

day 5 ~ 10

culture “spheroid” human mesoderm in “StemPro-34 medium” for “5” days. Medium should contain factors shown in the following table, and should be changed once in “5” days.

Ideally, “HE” should express “CD34”, “GJA4”, “GJA5”, “HEY2”, “DLL4”, “CXCR4” at the end of culture.

Step 7: endothelial 2

day 10 ~ 13

culture “adherent” human HE in “StemPro-34 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.