The miR-148/152 family contributes to angiogenesis of human pluripotent stem cell- derived endothelial cells by inhibiting MEOX2

DOI: 10.1016/j.omtn.2023.04.020

Authors: Fengyue Ding; Hongchun Wu; Xinglong Han; Xue Jiang; Yang Xiao; Yuanyuan Tu; Miao Yu; Wei Lei; Shijun Hu

Step 1: iPSC/ESC maintainance

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

culture “adherent” human ESC in “PSCeasy medium”. “ESC” should be “downregulated” for “miR-148/152”. For ECM, use “Matrigel” at “N/A” “N/A”.

Step 2: Primitive streak 1

day 1 ~ 3

culture “adherent” human ESC in “RPMI 1640” for “2” days. “ESC” should be “downregulated” for “miR-148/152”. 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.

Step 3: mesoderm 1

day 3 ~ 4

culture “adherent” human ESC in “RPMI 1640” for “1” days. “ESC” should be “downregulated” for “miR-148/152”. 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 4: endothelial 1

day 4 ~ 7

culture “adherent” human ESC in “RPMI 1640” for “3” days. “ESC” should be “downregulated” for “miR-148/152”. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “3” days.

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