GATA2 Is Dispensable for Specification of Hemogenic Endothelium but Promotes Endothelial-to-Hematopoietic Transition

DOI: 10.1016/j.stemcr.2018.05.002

Authors: HyunJun Kang; Walatta-Tseyon Mesquitta; Ho Sun Jung; Oleg V. Moskvin; James A. Thomson; Igor I. Slukvin

Step 1: iPSC/ESC maintainance

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

culture “adherent” human ESC in “E8”. “ESC” should be “downregulated” for “GATA2”. For ECM, use “Matrigel” at “N/A” “N/A”.

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

Step 2: Primitive streak 1

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

culture “adherent” human ESC in “E8”. “ESC” should be “downregulated” for “GATA2”. For ECM, use “Collagen” at “N/A” “N/A”.

Step 3: Primitive streak 2

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

culture “adherent” human ESC in “E8”. “ESC” should be “downregulated” for “GATA2”. For ECM, use “Collagen” at “N/A” “N/A”.

Step 4: mesoderm 1

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

culture “adherent” human ESC in “E8”. “ESC” should be “downregulated” for “GATA2”. For ECM, use “Collagen” at “N/A” “N/A”.

Step 5: mesoderm 2

day 4 ~ 5 (duration missing in database; assumed 1 day)

culture “adherent” human ESC in “E8”. “ESC” should be “downregulated” for “GATA2”. For ECM, use “Collagen” at “N/A” “N/A”.

Step 6: endothelial 1

day 5 ~ 6 (duration missing in database; assumed 1 day)

culture “adherent” human ESC in “E8”. “ESC” should be “downregulated” for “GATA2”. For ECM, use “Collagen” at “N/A” “N/A”.

Step 7: endothelial 2

day 6 ~ 12

culture “adherent” human EC in “unspecified medium” for “6” days. “EC” should be “downregulated” for “GATA2”. For ECM, use “Collagen” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “6” days.

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