Accelerated differentiation of human induced pluripotent stem cells to blood–brain barrier endothelial cells

DOI: 10.1186/s12987-017-0059-0

Authors: Emma K. Hollmann; Amanda K. Bailey; Archit V. Potharazu; M. Diana Neely; Aaron B. Bowman; Ethan S. Lippmann

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “E8” for “1” 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 “1” days.

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

Step 2: mesoderm 1

day 1 ~ 2

culture “adherent” human PSC in “E6 medium” for “1” days. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should be changed once in “1” days.

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

Step 3: endothelial 1

day 2 ~ 4

culture “adherent” human PSC in “Endothelial Serum-Free 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 express “PECAM1”, “GLUT-1”, “CDH5”, “occludin” at the end of culture.

Step 4: endothelial 2

day 4 ~ 5 (duration missing in database; assumed 1 day)

culture “device” human EC in “Endothelial Serum-Free Medium”. For ECM, use “collagen IV/fibronectin” at “N/A” “N/A”. Medium should contain factors shown in the following table.

Ideally, “EC” should express “GLUT-1”, “claudin-5”, “occludin”, “CDH5”, “PECAM1” at the end of culture.