Nox4 promotes endothelial differentiation through chromatin remodeling

DOI: 10.1016/j.redox.2022.102381

Authors: F. Hahner; F. Moll; T. Warwick; D.M. Hebchen; G.K. Buchmann; J. Epah; W. Abplanalp; T. Schader; S. Günther; R. Gilsbach; R.P. Brandes; K. Schröder

Step 1: iPSC/ESC maintainance

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

culture “adherent” mouse PSC in “DMEM”. “PSC” should be “downregulated” for “Nox4”. For ECM, use “feeder cells” at “N/A” “N/A”. Medium should contain factors shown in the following table.

Ideally, “PSC” should express “POU5F1”, “NANOG”, “SOX2”, “Ssea1”, “Klf4” at the end of culture.

Step 2: Primitive streak 1

day 1 ~ 5

culture “spheroid” mouse PSC in “alpha MEM” for “4” days. “PSC” should be “downregulated” for “Nox4”. Medium should contain factors shown in the following table, and should be changed once in “4” days.

Step 3: mesoderm 1

day 5 ~ 8

culture “spheroid” mouse PSC in “alpha MEM” for “3” days. “PSC” should be “downregulated” for “Nox4”. Medium should contain factors shown in the following table, and should be changed once in “3” days.

Ideally, “mesoderm” should express “MESP1”, “MIXL1”, “TBXT” at the end of culture.

Step 4: endothelial 1

day 8 ~ 10

culture “adherent” mouse PSC in “alpha MEM” for “2” days. “PSC” should be “downregulated” for “Nox4”. For ECM, use “Gelatin” at “0.2” “%”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Ideally, “EC” should express “CDH5”, “Nos3”, “PECAM1”, “Kdr” at the end of culture.