Inhibition of transforming growth factor beta signaling pathway promotes differentiation of human induced pluripotent stem cell-derived brain microvascular endothelial-like cells

DOI: 10.1186/s12987-020-00197-1

Authors: Misaki Yamashita; Hiromasa Aoki; Tadahiro Hashita; Takahiro Iwao; Tamihide Matsunaga

Step 1: iPSC/ESC maintainance

day 0 ~ 3

culture “adherent” human PSC in “StemSure hPSC 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, “PSC” should be obtained at the end of culture.

Step 2: endothelial 1

day 3 ~ 4

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

Step 3: endothelial 2

day 4 ~ 8

culture “adherent” human PSC in “Human Endothelial-SFM” for “4” days. For ECM, use “fibronectin/collagen IV” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “4” days.

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