SARS-CoV-2 infection of human pluripotent stem cell-derived vascular cells reveals smooth muscle cells as key mediators of vascular pathology during infection

DOI: 10.1038/s41467-024-54917-4

Authors: Alexsia Richards; Andrew S. Khalil; Max Friesen; Troy W. Whitfield; Xinlei Gao; Tenzin Lungjangwa; Roger D. Kamm; Zhengpeng Wan; Lee Gehrke; David Mooney; Rudolf Jaenisch

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.

Step 2: Primitive streak 1

day 1 ~ 2 (duration missing in database; assumed 1 day)

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

Step 3: mesoderm 1

day 2 ~ 3 (duration missing in database; assumed 1 day)

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

Step 4: endothelial 1

day 3 ~ 4 (duration missing in database; assumed 1 day)

culture “adherent” human PSC in “Vascular SFM”. For ECM, use “FN1” at “0.01” “g/l”. Medium should contain factors shown in the following table.

Ideally, “EC” should express “PECAM1”, “CDH5”, “VWF”, “ETV2”, “NRP1” at the end of culture.

Step 5: endothelial 2

day 4 ~ 5 (duration missing in database; assumed 1 day)

culture “adherent” human PSC in “Vascular SFM”. For ECM, use “FN1” at “0.01” “g/l”. Medium should contain factors shown in the following table.

Ideally, “EC” should express “PECAM1”, “CDH5”, “VWF”, “ETV2”, “NRP1” at the end of culture.