Exploring dysfunctional barrier phenotypes associated with glaucoma using a human pluripotent stem cell-based model of the neurovascular unit

DOI: 10.1186/s12987-024-00593-x

Authors: Sailee S. Lavekar; Jason M. Hughes; Cátia Gomes; Kang-Chieh Huang; Jade Harkin; Scott G. Canfield; Jason S. Meyer

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 be changed once in “1” days.

Step 2: endothelial 1

day 1 ~ 6

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 “Occludin”, “Claudin-5”, “ZO-1”, “PECAM” at the end of culture.

Step 3: endothelial 2

day 6 ~ 7 (duration missing in database; assumed 1 day)

culture “adherent” human PSC in “Endothelial Serum-Free Medium”. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table.