Wnt signaling controls the specification of definitive and primitive hematopoiesis from human pluripotent stem cells

DOI: 10.1038/nbt.2915

Authors: Christopher M Sturgeon; Andrea Ditadi; Geneve Awong; Marion Kennedy; Gordon Keller

Step 1: iPSC/ESC maintainance

day 0 ~ 1

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

Step 2: Primitive streak 1

day 1 ~ 4

culture “spheroid” human PSC in “SFD” for “3” days. Medium should contain factors shown in the following table, and should be changed once in “3” days.

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

Step 3: mesoderm 1

day 4 ~ 5

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

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

Step 4: mesoderm 2

day 5 ~ 11

culture “spheroid” human PSC in “StemPro” for “6” days. Medium should contain factors shown in the following table, and should be changed once in “6” days.

Ideally, “EC” should express “CD34” at the end of culture.

Step 5: endothelial 1

day 11 ~ 18

culture “adherent” unknown species cell in “StemPro” for “7” 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 “7” days.

Ideally, “EC” should express “CDH5” at the end of culture. Ideally, “hematopoietic cells” should express “PTPRC” at the end of culture.