Reactive astrocytes transduce inflammation in a blood-brain barrier model through a TNF-STAT3 signaling axis and secretion of alpha 1-antichymotrypsin

DOI: 10.1038/s41467-022-34412-4

Authors: Hyosung Kim; Kun Leng; Jinhee Park; Alexander G. Sorets; Suil Kim; Alena Shostak; Rebecca J. Embalabala; Kate Mlouk; Ketaki A. Katdare; Indigo V. L. Rose; Sarah M. Sturgeon; Emma H. Neal; Yan Ao; Shinong Wang; Michael V. Sofroniew; Jonathan M. Brunger; Douglas G. McMahon; Matthew S. Schrag; Martin Kampmann; Ethan S. Lippmann

Step 1: iPSC/ESC maintainance

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

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

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

Step 2: endothelial 1

day 1 ~ 2

culture “adherent” human PSC in “E6 medium” for “1” days. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should be changed once in “1” days.

Ideally, “EC” should express “CDH5”, “TEK”, “VWF”, “SLC2A1” at the end of culture.

Step 3: endothelial 2

day 2 ~ 4

culture “adherent” human PSC in “unspecified 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.