Intrinsic epigenetic control of angiogenesis in induced pluripotent stem cell-derived endothelium regulates vascular regeneration

DOI: 10.1038/s41536-022-00223-w

Authors: Bria L. Macklin; Ying-Yu Lin; Kevin Emmerich; Emily Wisniewski; Brian M. Polster; Konstantinos Konstantopoulos; Jeff S. Mumm; Sharon Gerecht

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “E8” for “1” days. For ECM, use “Vitronectin” at “N/A” “N/A”. Medium 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 “Vitronectin” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Step 3: mesoderm 1

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

culture “unspecified method” human PSC in “alpha MEM”.

Step 4: endothelial 1

day 3 ~ 5

culture “adherent” human PSC in “ECGM” for “2” days. For ECM, use “COL1A1” 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 express “KDR”, “HK1”, “HK2”, “PFKFB3”, “VEGF” at the end of culture.