Derivation of blood-brain barrier endothelial cells from human pluripotent stem cells

DOI: 10.1038/nbt.2247

Authors: Ethan S Lippmann; Samira M Azarin; Jennifer E Kay; Randy A Nessler; Hannah K Wilson; Abraham Al-Ahmad; Sean P Palecek; Eric V Shusta

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 be obtained at the end of culture.

Step 2: endothelial 1

day 2 ~ 7

culture “adherent” human PSC in “DMEM/F12” for “5” 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 “5” days.

Ideally, “EC” should express “PECAM1”, “claudin-5”, “occludin”, “p-glycoprotein”, “GLUT-1” at the end of culture.

Step 3: endothelial 2

day 7 ~ 9

culture “adherent” human PSC in “Endothelial Serum-Free Medium” for “2” 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 “2” days.

Ideally, “EC” should express “PECAM1”, “claudin-5”, “occludin”, “p-glycoprotein”, “GLUT-1”, “VWF”, “CDH5”, “ZO-1”, “LDLR”, “INSR”, “LEPR”, “BCAM”, “TFRC”, “AGER”, “SLC7A5”, “SLC1A1”, “SLC38A5”, “SLC16A1”, “ABCG2”, “ABCC1”, “ABCC2”, “ABCC4”, “STRA6”, “SLC2A1”, “ABCB1” at the end of culture.