ZEB2 and MEIS1 independently contribute to hematopoiesis via early hematopoietic enhancer activation

DOI: 10.1016/j.isci.2023.107893

Authors: Yohko Kitagawa; Akihiro Ikenaka; Ryohichi Sugimura; Akira Niwa; Megumu K. Saito

Step 1: iPSC/ESC maintainance

day 0 ~ 3

culture “adherent” human PSC in “mTeSR” for “3” days. For ECM, use “Laminin” at “N/A” “N/A”. Medium should be changed once in “3” days.

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

Step 2: Primitive streak 1

day 3 ~ 5

culture “adherent” human PSC in “E8” for “2” days. For ECM, use “Laminin” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Step 3: mesoderm 1

day 5 ~ 7

culture “adherent” human PSC in “E6 medium” for “2” days. For ECM, use “Laminin” at “N/A” “N/A”. 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: endothelial 1

day 7 ~ 9

culture “adherent” human PSC in “E6 medium” for “2” days. For ECM, use “Laminin” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

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

Step 5: endothelial 2

day 9 ~ 12

culture “adherent” human PSC in “StemLine II” for “3” days. For ECM, use “Retronectin” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “3” days.

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