P450 oxidoreductase regulates barrier maturation by mediating retinoic acid metabolism in a model of the human BBB

DOI: 10.1016/j.stemcr.2022.07.010

Authors: Dor Zlotnik; Tatiana Rabinski; Aviv Halfon; Shira Anzi; Inbar Plaschkes; Hadar Benyamini; Yuval Nevo; Orly Yahalom Gershoni; Benyamin Rosental; Eli Hershkovitz; Ayal Ben-Zvi; Gad D. Vatine

Step 1: iPSC/ESC maintainance

day 0 ~ 3

culture “unspecified method” human PSC in “NutriStem” for “3” days. Medium should be changed once in “3” days.

Step 2: Primitive streak 1

day 3 ~ 4 (duration missing in database; assumed 1 day)

culture “unspecified method” human PSC in “DMEM/F12”. Medium should contain factors shown in the following table.

Step 3: endothelial 1

day 4 ~ 6

culture “adherent” human PSC in “Endothelial SFM” for “2” days. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Ideally, “EC” should express “ZO-1”, “GLUT-1”, “CLDN-5”, “OCLN” at the end of culture.

Step 4: endothelial 2

day 6 ~ 7 (duration missing in database; assumed 1 day)

culture “adherent” unknown species cell in “Endothelial SFM”. For ECM, use “fibronectin + collagen IV” at “N/A” “N/A”.