Differentiation of human pluripotent stem cells to brain microvascular endothelial cell-like cells suitable to study immune cell interactions

DOI: 10.1016/j.xpro.2021.100563

Authors: Hideaki Nishihara; Benjamin D. Gastfriend; Pelin Kasap; Sean P. Palecek; Eric V. Shusta; Britta Engelhardt

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “mTeSR” 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: mesoderm 1

day 1 ~ 2

culture “adherent” human PSC in “LaSR basal medium” 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 3: mesoderm 2

day 2 ~ 3

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

Ideally, “EPC” should express “PECAM1” at the end of culture.

Step 4: endothelial 1

day 3 ~ 6

culture “adherent” human EPC in “Human Endothelial SFM” for “3” days. For ECM, use “Collagen IV” at “0.01” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “3” days.