PU.1 drives specification of pluripotent stem cell-derived endothelial cells to LSEC-like cells

DOI: 10.1038/s41419-020-03356-2

Authors: Jonathan De Smedt; Elise Anne van Os; Irene Talon; Sreya Ghosh; Burak Toprakhisar; Rodrigo Furtado Madeiro Da Costa; Samantha Zaunz; Marta Aguirre Vazquez; Ruben Boon; Pieter Baatsen; Ayla Smout; Stefaan Verhulst; Leo A. van Grunsven; Catherine M. Verfaillie

Step 1: iPSC/ESC maintainance

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

culture “adherent” human ESC in “E8 Flex medium”. Medium should contain factors shown in the following table.

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

Step 2: endothelial 1

day 1 ~ 13

culture “adherent” human ESC in “unspecified medium” for “12” days. “ESC” should be “upregulated” for “ETV2”. Medium should contain factors shown in the following table, and should be changed once in “12” days.

Ideally, “EC” should express “CDH5”, “PECAM1”, “KDR”, “FLI1”, “ERG” at the end of culture.

Step 3: endothelial 2

day 13 ~ 25

culture “adherent” human ESC in “unspecified medium” for “12” days. “ESC” should be “upregulated” for “SPI1”. Medium should contain factors shown in the following table, and should be changed once in “12” days.

Ideally, “EC” should express “FCGR2B”, “LYVE1”, “MRC1”, “STAB1”, “STAB2”, “FCN3”, “OIT3”, “CRHBP” at the end of culture.