Role of Notch1 in the arterial specification and angiogenic potential of mouse embryonic stem cell-derived endothelial cells

DOI: 10.1186/s13287-018-0945-7

Authors: Jae Kyung Park; Tae Wook Lee; Eun Kyoung Do; Hye Ji Moon; Jae Ho Kim

Step 1: iPSC/ESC maintainance

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

culture “adherent” mouse ESC in “DMEM”. For ECM, use “Fibroblast” at “N/A” “N/A”. Medium should contain factors shown in the following table.

Ideally, “ESC” should express “POU5F1”, “SSEA-1”, “SOX2”, “NANOG” at the end of culture.

Step 2: mesoderm 1

day 1 ~ 6

culture “adherent” mouse ESC in “IMDM” for “5” days. For ECM, use “Collagen IV” at “0.01” “g/m2”. Medium should contain factors shown in the following table, and should be changed once in “5” days.

Ideally, “mesoderm” should express “KDR” at the end of culture.

Step 3: endothelial 1

day 6 ~ 9

culture “adherent” mouse mesoderm in “IMDM” for “3” days. “mesoderm” should be “upregulated” for “NOTCH1”. For ECM, use “Collagen IV” at “0.01” “g/m2”. Medium should contain factors shown in the following table, and should be changed once in “3” days.

Ideally, “EC” should express “PECAM1”, “CDH5”, “CXCR4”, “NRP1”, “Ephrin B2”, “HEY1”, “Hey2”, “HES1”, “Foxc2”, “Jag1”, “DLL4” at the end of culture.