Endothelial progenitor cells and neural progenitor cells synergistically protect cerebral endothelial cells from Hypoxia/reoxygenation-induced injury via activating the PI3K/Akt pathway

DOI: 10.1186/s13041-016-0193-7

Authors: Jinju Wang; Yusen Chen; Yi Yang; Xiang Xiao; Shuzhen Chen; Cheng Zhang; Bradley Jacobs; Bin Zhao; Ji Bihl; Yanfang Chen

Step 1: iPSC/ESC maintainance

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

culture “adherent” human PSC in “mTeSR”. For ECM, use “Matrigel” at “N/A” “N/A”.

Ideally, “PSC” should express “SOX2”, “POU5F1” at the end of culture.

Step 2: Primitive streak 1

day 1 ~ 6

culture “spheroid” human PSC in “DMEM/F12” for “5” days. Medium should contain factors shown in the following table, and should be changed once in “5” days.

Step 3: endothelial 1

day 6 ~ 13

culture “adherent” human PSC in “EBM-2 medium” for “7” days. For ECM, use “Gelatin” at “0.1” “%”. Medium should contain factors shown in the following table, and should be changed once in “7” days.

Ideally, “EPC” should express “CD34”, “KDR” at the end of culture.