A human pluripotent stem cell-derived in vitro model of the blood–brain barrier in cerebral malaria

DOI: 10.1186/s12987-024-00541-9

Authors: Adnan Gopinadhan; Jason M. Hughes; Andrea L. Conroy; Chandy C. John; Scott G. Canfield; Dibyadyuti Datta

Step 1: iPSC/ESC maintainance

day 0 ~ 3

culture “adherent” human PSC in “mTeSR” for “3” days. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should be changed once in “3” days.

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

Step 2: Primitive streak 1

day 3 ~ 4

culture “adherent” human PSC in “DMEM/F12” 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.

Step 3: endothelial 1

day 4 ~ 6

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 be obtained at the end of culture.

Step 4: endothelial 2

day 6 ~ 7 (duration missing in database; assumed 1 day)

culture “adherent” human PSC in “Endothelial Serum-Free Medium”. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table.

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