FLI1 and PKC co-activation promote highly efficient differentiation of human embryonic stem cells into endothelial-like cells

DOI: 10.1038/s41419-017-0162-9

Authors: Hao Zhao; Yan Zhao; Zili Li; Qi Ouyang; Yi Sun; Di Zhou; Pingyuan Xie; Sicong Zeng; Lingfeng Dong; Hua Wen; Guangxiu Lu; Ge Lin; Liang Hu

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “mTeSR” for “1” days. “PSC” should be “unspecified regulation” for “FLI1”. 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.

Step 2: endothelial 1

day 1 ~ 2

culture “adherent” human PSC in “mTeSR” for “1” days. “PSC” should be “upregulated” for “FLI1”. 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”, “CDH5”, “KDR”, “VWF”, “FLT1”, “CD34”, “NRP2”, “NOTCH1”, “TGFB1”, “VEGF”, “GATA2”, “ERG”, “MEIS1”, “POU5F1”, “SOX2”, “NANOG”, “UTF1”, “C-MYC” at the end of culture.

Step 3: endothelial 2

day 2 ~ 3

culture “adherent” human PSC in “mTeSR” for “1” days. “PSC” should be “unspecified regulation” for “FLI1”. 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.