A Simplified, Fully Defined Differentiation Scheme for Producing Blood-Brain Barrier Endothelial Cells from Human iPSCs

DOI: 10.1016/j.stemcr.2019.05.008

Authors: Emma H. Neal; Nicholas A. Marinelli; Yajuan Shi; P. Mason McClatchey; Kylie M. Balotin; Dalton R. Gullett; Kameron A. Hagerla; Aaron B. Bowman; Kevin C. Ess; John P. Wikswo; Ethan S. Lippmann

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “E8” for “1” 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 “1” days.

Step 2: Primitive streak 1

day 1 ~ 2

culture “adherent” human PSC in “E6 medium” for “1” days. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should be changed once in “1” days.

Step 3: endothelial 1

day 2 ~ 4

culture “adherent” human PSC in “hESFM” 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 “occludin”, “claudin-5”, “CDH5”, “GLUT-1”, “PECAM1”, “P-glycoprotein”, “MRP” at the end of culture.

Step 4: endothelial 2

day 4 ~ 5

culture “adherent” human PSC in “hESFM” for “1” days. For ECM, use “collagen IV/fibronectin” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “1” days.