Presence of a mutation in PSEN1 or PSEN2 gene is associated with an impaired brain endothelial cell phenotype in vitro

DOI: 10.1186/s12987-020-00235-y

Authors: Snehal Raut; Ronak Patel; Abraham J. Al-Ahmad

Step 1: iPSC/ESC maintainance

day 0 ~ 5

culture “adherent” unknown species cell and 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: mesoderm 1

day 5 ~ 11

culture “adherent” unknown species cell and human PSC in “DMEM/F12” for “6” 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 “6” days.

Step 3: endothelial 1

day 11 ~ 13

culture “adherent” unknown species cell and human PSC in “EC medium” 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 be obtained at the end of culture.

Step 4: endothelial 2

day 13 ~ 14

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

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