Laminin 221 fragment is suitable for the differentiation of human induced pluripotent stem cells into brain microvascular endothelial-like cells with robust barrier integrity

DOI: 10.1186/s12987-020-00186-4

Authors: Hiromasa Aoki; Misaki Yamashita; 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 “LN221F” at “0.01” “g/m2”. Medium should contain factors shown in the following table, and should be changed once in “3” days.

Ideally, “PSC” should express “POU5F1”, “NANOG” 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 “LN221F” at “0.01” “g/m2”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Step 3: endothelial 2

day 4 ~ 7

culture “device” human PSC in “HE-SFM” for “3” days. For ECM, use “LN221F” at “0.1” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “3” days.

Ideally, “EC” should express “CDH5”, “Claudin-5”, “MDR1”, “BCRP”, “Occludin”, “ZO-1”, “GLUT1”, “LAT1”, “PECAM1”, “VWF”, “KDR” at the end of culture.