Therapeutic correction of hemophilia A by transplantation of hPSC-derived liver sinusoidal endothelial cell progenitors

DOI: 10.1016/j.celrep.2022.110621

Authors: Blair K. Gage; Simone Merlin; Cristina Olgasi; Antonia Follenzi; Gordon M. Keller

Step 1: iPSC/ESC maintainance

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

culture “adherent” human PSC in “DMEM/F12”. For ECM, use “Matrigel” at “0.25” “%”. Medium should contain factors shown in the following table.

Step 2: mesoderm 1

day 1 ~ 4

culture “spheroid” human PSC in “IMDM” for “3” days. For ECM, use “PolyHema” at “5” “% w/v”. Medium should contain factors shown in the following table, and should be changed once in “3” days.

Ideally, “mesoderm” should express “KDR”, “CD235a”, “CD235b”, “CD34”, “ETV2” at the end of culture.

Step 3: endothelial 1

day 4 ~ 6

culture “adherent” human PSC in “IMDM” for “2” days. For ECM, use “Matrigel” at “2.5” “%”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Ideally, “EC” should express “CD34”, “PECAM1”, “CD73”, “NR2F2”, “NRP2”, “NT5E”, “EPHB4” at the end of culture.