Induced pluripotent stem cells with NOTCH1 gene mutation show impaired differentiation into smooth muscle and endothelial cells: Implications for bicuspid aortic valve-related aortopathy

DOI: 10.1016/j.jtcvs.2018.02.087

Authors: Jiao Jiao; Weihua Tian; Ping Qiu; Elizabeth L. Norton; Michael M. Wang; Y. Eugene Chen; Bo Yang

Step 1: iPSC/ESC maintainance

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

culture “adherent” human PSC in “E8”. “PSC” should be “downregulated” for “NOTCH1”. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table.

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

Step 2: mesoderm 1

day 1 ~ 2

culture “adherent” human PSC in “DMEM/F12” for “1” days. “PSC” should be “downregulated” for “NOTCH1”. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Ideally, “CVPC” should express “ISL1”, “NKX2.5”, “myocardin” at the end of culture.

Step 3: endothelial 1

day 2 ~ 4

culture “adherent” human CVPC in “Stempro34” for “2” days. “CVPC” should be “downregulated” for “NOTCH1”. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Ideally, “EC” should express “PECAM1”, “CD105” at the end of culture.