Cytokine-free directed differentiation of human pluripotent stem cells efficiently produces hemogenic endothelium with lymphoid potential

DOI: 10.1186/s13287-017-0519-0

Authors: Yekaterina Galat; Svetlana Dambaeva; Irina Elcheva; Aaruni Khanolkar; Kenneth Beaman; Philip M. Iannaccone; Vasiliy Galat

Step 1: iPSC/ESC maintainance

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

culture “adherent” human PSC in “mTeSR”. For ECM, use “Matrigel” at “N/A” “N/A”.

Ideally, “PSC” should express “AP”, “TRA-1-80”, “NANOG”, “TRA-1-60”, “POU5F1” at the end of culture.

Step 2: Primitive streak 1

day 1 ~ 3

culture “adherent” human PSC in “DMEM/F12” for “2” 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 “2” days.

Step 3: endothelial 1

day 3 ~ 6

culture “adherent” human PSC in “DMEM/F12” 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.

Ideally, “HE” should express “CD34”, “PECAM1” at the end of culture.

Step 4: endothelial 2

day 6 ~ 7 (duration missing in database; assumed 1 day)

culture “adherent” human PSC in “Vasculife medium”.

Ideally, “EC” should express “VWF”, “PECAM1”, “CDH5”, “CDH5” at the end of culture.