VGLL4 promotes vascular endothelium specification via TEAD1 in the vascular organoids and human pluripotent stem cells-derived endothelium model

DOI: 10.1007/s00018-023-04858-w

Authors: Yingyi Quan; Minjie Hu; Jiang Jiang; Peifeng Jin; Junming Fan; Ming Li; Xiaofang Fan; Yongsheng Gong; Yang Yang; Yongyu Wang

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human ESC in “Nuwacell hiPSC/hESC medium-ncEpic” for “1” days. 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, “ESC” should be obtained at the end of culture.

Step 2: mesoderm 1

day 1 ~ 4

culture “adherent” human ESC in “N2B27 medium” for “3” days. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “3” days.

Ideally, “ME” should express “TBXT”, “EOMES”, “MIXL1” at the end of culture.

Step 3: endothelial 1

day 4 ~ 5

culture “adherent” human ME in “StemPro” for “1” days. “ME” should be “upregulated” for “VGLL4”. 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, “EC” should express “PECAM1”, “VE-CAD”, “VWF”, “VGLL4”, “FLI1”, “ERG”, “FOXC2”, “SOX17”, “HEY1”, “TEAD1”, “TEAD4” at the end of culture.

Step 4: endothelial 2

day 5 ~ 6 (duration missing in database; assumed 1 day)

culture “unspecified method” human EC in “EGM2”.

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