Brain microvascular endothelial cell dysfunction in an isogenic juvenile iPSC model of Huntington’s disease

DOI: 10.1186/s12987-022-00347-7

Authors: Raleigh M. Linville; Renée F. Nerenberg; Gabrielle Grifno; Diego Arevalo; Zhaobin Guo; Peter C. Searson

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “mTeSR” for “1” days. For ECM, use “Matrigel” at “0.083” “g/l”. Medium should be changed once in “1” days.

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

Step 2: mesoderm 1

day 1 ~ 2

culture “adherent” human PSC in “DMEM/F12” for “1” days. For ECM, use “Matrigel” at “0.083” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Step 3: endothelial 1

day 2 ~ 3

culture “adherent” human PSC in “Endothelial cell serum-free medium” for “1” days. For ECM, use “Matrigel” at “0.083” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Ideally, “EC” should express “PECAM1”, “GLUT-1” at the end of culture.

Step 4: endothelial 2

day 3 ~ 4

culture “adherent” human EC in “Endothelial cell serum-free medium” for “1” days. For ECM, use “collagen IV + fibronectin” at “0.05” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Ideally, “EC” should express “PECAM1”, “GLUT-1”, “claudin-5”, “CDH5”, “ZO1”, “occludin”, “P-glycoprotein”, “KDR”, “IL4R”, “CCR1”, “CXCR4”, “IL17RD”, “CXCL12” at the end of culture.