A Perfused In Vitro Human iPSC-Derived Blood–Brain Barrier Faithfully Mimics Transferrin Receptor-Mediated Transcytosis of Therapeutic Antibodies

DOI: 10.1007/s10571-023-01404-x

Authors: Floriana Burgio; Carine Gaiser; Kevin Brady; Viviana Gatta; Reiner Class; Ramona Schrage; Laura Suter-Dick

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 “0.333” “g/l”.

Ideally, “PSC” should express “POU5F1”, “SSEA4”, “TRA-1-60”, “SOX2”, “ZO1”, “Occludin”, “transferrin receptor”, “P-gp” at the end of culture.

Step 2: endothelial 1

day 1 ~ 6

culture “adherent” human PSC in “DMEM/F12” for “5” days. For ECM, use “Matrigel” at “0.333” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “5” days.

Step 3: endothelial 2

day 6 ~ 8

culture “adherent” human PSC in “Endothelial cell serum-free medium” for “2” days. For ECM, use “Matrigel” at “0.333” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Ideally, “EC” should express “ABCB1”, “ABCC1”, “ABCG2”, “SLC2A1”, “TRFC”, “ZO1”, “Occludin”, “Claudin5”, “Glut1”, “insulin receptor”, “LDL receptor”, “PECAM1”, “VWF”, “P-gp” at the end of culture.