Derivation of Brain Capillary-like Endothelial Cells from Human Pluripotent Stem Cell-Derived Endothelial Progenitor Cells

DOI: 10.1016/j.stemcr.2019.08.002

Authors: Catarina Praça; Susana C. Rosa; Emmanuel Sevin; Romeo Cecchelli; Marie-Pierre Dehouck; Lino S. Ferreira

Step 1: iPSC/ESC maintainance

day 0 ~ 1 (duration missing in database; assumed 1 day)

culture “adherent” human PSC in “unspecified medium”. For ECM, use “FN1” at “0.01” “g/m2”. Medium should contain factors shown in the following table.

Step 2: mesoderm 1

day 1 ~ 11

culture “adherent” unknown species PSC in “unspecified medium” for “10” days. For ECM, use “FN1” at “0.01” “g/m2”. Medium should be changed once in “10” days.

Ideally, “EPC” should express “PECAM1”, “KDR”, “GLUT-1”, “CLAUDIN-5”, “OCCLUDIN” at the end of culture.

Step 3: mesoderm 2

day 11 ~ 12 (duration missing in database; assumed 1 day)

culture “adherent” human EPC in “EGM2”. For ECM, use “FN1” at “0.01” “g/m2”. Medium should contain factors shown in the following table.

Step 4: endothelial 1

day 12 ~ 27

culture “adherent” unknown species EPC in “EGM2” for “15” days. For ECM, use “FN1” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “15” days.

Ideally, “EC” should express “VE-CADHERIN”, “CLAUDIN-5”, “ZO-1”, “OCCLUDIN”, “GLUT-1”, “PECAM1”, “TIE-2”, “VWF”, “CLDN1”, “ZO1”, “SLC16A1”, “INSR”, “TFRC”, “LRP1”, “ABCB1”, “ABCC1”, “ABCC5”, “PGP” at the end of culture.