SREBP1 suppresses the differentiation and epithelial function of hiPSC-derived endothelial cells by inhibiting the microRNA199b-5p pathway

DOI: 10.1016/j.scr.2021.102174

Authors: Xin Qian; Xiangjiang Guo; Qihong Ni; Han Wang; Meng Ye; Lan Zhang

Step 1: iPSC/ESC maintainance

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

culture “adherent” human PSC in “mTeSR”. For ECM, use “Matrigel” at “N/A” “N/A”.

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

Step 2: Primitive streak 1

day 1 ~ 4

culture “adherent” human PSC in “DMEM/F12” 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.

Step 3: endothelial 1

day 4 ~ 7

culture “adherent” human PSC in “Human Endothelial Serum-free 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.

Step 4: endothelial 2

day 7 ~ 8 (duration missing in database; assumed 1 day)

culture “adherent” human EC in “Human Endothelial Serum-free Medium”. For ECM, use “Gelatin” at “0.1” “%”. Medium should contain factors shown in the following table.

Ideally, “EC” should express “PECAM1”, “KDR”, “CD34” at the end of culture.