Human isogenic cells of the neurovascular unit exert transcriptomic cell type-specific effects on a blood-brain barrier in vitro model of late-onset Alzheimer disease

DOI: 10.1186/s12987-023-00471-y

Authors: Undine Haferkamp; Carla Hartmann; Chaudhry Luqman Abid; Andreas Brachner; Alevtina Höchner; Anna Gerhartl; Bernadette Harwardt; Selin Leckzik; Jennifer Leu; Marco Metzger; Marina Nastainczyk-Wulf; Winfried Neuhaus; Sabrina Oerter; Ole Pless; Dan Rujescu; Matthias Jung; Antje Appelt-Menzel

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “mTeSR” for “1” days. For ECM, use “Matrigel” at “8.3e-5” “g/well”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

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

Step 2: Primitive streak 1

day 1 ~ 7

culture “adherent” human PSC in “DMEM/F12” for “6” days. For ECM, use “Matrigel” at “8.3e-5” “g/well”. Medium should contain factors shown in the following table, and should be changed once in “6” days.

Step 3: endothelial 1

day 7 ~ 9

culture “adherent” human PSC in “human endothelial serum-free medium” for “2” days. For ECM, use “Matrigel” at “8.3e-5” “g/well”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Step 4: endothelial 2

day 9 ~ 10

culture “device” human PSC in “human endothelial serum-free medium” for “1” days. For ECM, use “Collagen” at “1” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Ideally, “EC” should express “SLC2A1”, “TJP1”, “OCLN”, “CLDN5” at the end of culture.