Evaluation of a human iPSC-derived BBB model for repeated dose toxicity testing with cyclosporine A as model compound

DOI: 10.1016/j.tiv.2021.105112

Authors: Sara Wellens; Lucie Dehouck; Vidya Chandrasekaran; Pranika Singh; Rodrigo Azevedo Loiola; Emmanuel Sevin; Thomas Exner; Paul Jennings; Fabien Gosselet; Maxime Culot

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “mTeSR” for “1” days. For ECM, use “Matrigel” at “0.083” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Step 2: Primitive streak 1

day 1 ~ 2

culture “adherent” human PSC in “DMEM/F12” for “1” days. For ECM, use “Matrigel” at “0.083” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Step 3: mesoderm 1

day 2 ~ 3

culture “adherent” human PSC in “DMEM/F12” for “1” days. For ECM, use “Matrigel” at “0.083” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Step 4: endothelial 1

day 3 ~ 5

culture “adherent” human PSC in “human endothelial serum-free medium” for “2” days. For ECM, use “Matrigel” at “0.083” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Step 5: endothelial 2

day 5 ~ 7

culture “adherent” human PSC in “human endothelial serum-free medium” for “2” days. For ECM, use “Matrigel” at “0.1” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Ideally, “EC” should express “CLDN5”, “CDH5”, “ABCB1”, “ABCG2” at the end of culture.