Human forebrain endothelial cell therapy for psychiatric disorders

DOI: 10.1038/s41380-020-0839-9

Authors: Debkanya Datta; Sivan Subburaju; Sarah Kaye; Jugajyoti Baruah; Yong Kee Choi; Yeqi Nian; Jahan S. Khalili; Sangmi Chung; Abdallah Elkhal; Anju Vasudevan

Step 1: iPSC/ESC maintainance

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

culture “adherent” human ESC in “E8”. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table.

Step 2: mesoderm 1

day 1 ~ 3

culture “adherent” human ESC in “E8” 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.

Step 3: endothelial 1

day 3 ~ 6

culture “adherent” human ESC in “E6 medium” for “3” 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 “3” days.

Ideally, “EC” should express “PECAM1”, “CDH5”, “VWF”, “GABRB3”, “GABA”, “NKX2.1”, “CXCR4” at the end of culture.

Step 4: endothelial 2

day 6 ~ 8

culture “adherent” human ESC in “E6 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”, “VWF”, “GABRB3”, “CDH5”, “NKX2.1”, “PAX6”, “ISL1”, “GABA”, “CXCR4” at the end of culture.