Growth-factor reduced Matrigel source influences stem cell derived brain microvascular endothelial cell barrier properties

DOI: 10.1186/s12987-016-0030-5

Authors: Ronak Patel; Abraham J. Alahmad

Step 1: iPSC/ESC maintainance

day 0 ~ 5

culture “adherent” human PSC in “E8” for “5” days. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should be changed once in “5” days.

Ideally, “PSC” should be obtained at the end of culture.

Step 2: endothelial 1

day 5 ~ 6

culture “adherent” human PSC in “DMEM/F12” for “1” days. For ECM, use “T-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: endothelial 2

day 6 ~ 8

culture “adherent” human PSC in “EC SFM” for “2” days. For ECM, use “T-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 “PECAM1”, “CDH5”, “SLC2A1”, “CLDN5”, “OCLN”, “ABCB1”, “ABCG2”, “ABCC1” at the end of culture.