Nox2 contributes to the arterial endothelial specification of mouse induced pluripotent stem cells by upregulating Notch signaling

DOI: 10.1038/srep33737

Authors: Xueling Kang; Xiangxiang Wei; Xinhong Wang; Li Jiang; Cong Niu; Jianyi Zhang; Sifeng Chen; Dan Meng

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 “Nox2”. Medium should contain factors shown in the following table.

Ideally, “PSC” should express “POU5F1”, “SSEA-1” at the end of culture.

Step 2: Primitive streak 1

day 1 ~ 5

culture “spheroid” mouse PSC in “DMEM” for “4” days. “PSC” should be “downregulated” for “Nox2”. Medium should contain factors shown in the following table, and should be changed once in “4” days.

Step 3: endothelial 1

day 5 ~ 7

culture “adherent” mouse PSC in “DMEM” for “2” days. “PSC” should be “downregulated” for “Nox2”. For ECM, use “Gelatin” at “0.2” “%”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Step 4: endothelial 2

day 7 ~ 9

culture “adherent” mouse PSC in “DMEM” for “2” days. “PSC” should be “downregulated” for “Nox2”. 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 “PECAM1”, “CDH5”, “VWF”, “NOS3”, “EphrinB2”, “ALK1”, “Nrp1”, “JAG1”, “DLL4”, “NOTCH1”, “HES1”, “HEY1”, “VEGF”, “Ang-1”, “KDR”, “Tie-2”, “Ang-2”, “Nox2”, “Nox4” at the end of culture. Ideally, “EC” should express “EphrinB2”, “ALK1”, “Nrp1”, “JAG1”, “DLL4”, “NOTCH1”, “HES1”, “HEY1”, “Nox2” at the end of culture.