Development of Human in vitro Brain-blood Barrier Model from Induced Pluripotent Stem Cell-derived Endothelial Cells to Predict the in vivo Permeability of Drugs

DOI: 10.1007/s12264-019-00384-7

Authors: Yuan Li; Xueying Sun; Houfu Liu; Liang Huang; Guofeng Meng; Yu Ding; Wenji Su; Jiaqi Lu; Sophie Gong; Georg C. Terstappen; Ru Zhang; Wandong Zhang

Step 1: iPSC/ESC maintainance

day 0 ~ 2

culture “adherent” human PSC in “mTeSR” for “2” days. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should be changed once in “2” days.

Ideally, “PSC” should express “POU5F1”, “TRA1-60” at the end of culture.

Step 2: Primitive streak 1

day 2 ~ 8

culture “adherent” human PSC in “DMEM/F12” for “6” 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 “6” days.

Step 3: endothelial 1

day 8 ~ 10

culture “adherent” human PSC in “Human Endothelial-SFM” for “2” 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 “2” days.

Ideally, “EC” should express “VWF”, “Ulex”, “TJP1”, “CLDN5”, “OCLN”, “IGF1R”, “SLC3A2”, “SLC2A1”, “LRP1”, “TfR”, “ABCB1”, “ABCC1”, “ABCG2” at the end of culture.

Step 4: endothelial 2

day 10 ~ 13

culture “adherent” human EC in “Human Endothelial-SFM” for “3” days. For ECM, use “collagen IV, fibronectin” at “0.4” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “3” days.

Ideally, “EC” should express “VWF”, “Ulex”, “TJP1”, “CLDN5”, “OCLN”, “IGF1R”, “SLC3A2”, “SLC2A1”, “LRP1”, “TfR”, “ABCB1”, “ABCC1”, “ABCG2” at the end of culture.