Dual effect of TAT functionalized DHAH lipid nanoparticles with neurotrophic factors in human BBB and microglia cultures

DOI: 10.1186/s12987-022-00315-1

Authors: Sara Hernando; Polyxeni Nikolakopoulou; Dimitrios Voulgaris; Rosa Maria Hernandez; Manoli Igartua; Anna Herland

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “mTeSR” for “1” days. For ECM, use “Matrigel” at “8.3e-5” “g/well plate”. 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: Primitive streak 1

day 1 ~ 2

culture “adherent” human PSC in “E6 medium” for “1” days. For ECM, use “Matrigel” at “8.3e-5” “g/well plate”. Medium should be changed once in “1” days.

Step 3: endothelial 1

day 2 ~ 4

culture “adherent” human PSC in “hESFM” for “2” days. For ECM, use “Matrigel” at “8.3e-5” “g/well plate”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Step 4: endothelial 2

day 4 ~ 5

culture “device” human PSC in “hESFM” for “1” days. For ECM, use “collagen IV + fibronectin” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Ideally, “EC” should express “ZO-1” at the end of culture.