Inhibition of the E3 ligase UBR5 stabilizes TERT and protects vascular organoids from oxidative stress

DOI: 10.1186/s12967-024-05887-0

Authors: Haijing Zhao; Nian Cao; Qi Liu; Yingyue Zhang; Rui Jin; Huiying Lai; Li Zheng; Honghong Zhang; Yue Zhu; Yuhan Ma; Zengao Yang; Zhengfeng Wu; Weini Li; Yuqi Liu; Long Cheng; Yundai Chen

Step 1: iPSC/ESC maintainance

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

culture “unspecified method” human PSC and human PSC in “E8”. “PSC” should be “downregulated” for “UBR5”. “PSC” should be “upregulated” for “TERT”.

Step 2: Primitive streak 1

day 1 ~ 4

culture “spheroid” human PSC in “N2B27 medium” for “3” days. Medium should contain factors shown in the following table, and should be changed once in “3” days.

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

Step 3: mesoderm 1

day 4 ~ 6

culture “spheroid” human PSC in “N2B27 medium” for “2” days. Medium should contain factors shown in the following table, and should be changed once in “2” days.

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

Step 4: endothelial 1

day 6 ~ 8

culture “spheroid” human PSC in “StemPro” for “2” days. For ECM, use “Collagen I:Matrigel (1:1)” 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 “PECAM1” at the end of culture.

Step 5: endothelial 2

day 8 ~ 10

culture “spheroid” human PSC in “StemPro” for “2” days. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Ideally, “EC” should express “PECAM1” at the end of culture. Ideally, “Pericyte/SMC” should express “PDGFR-β” at the end of culture. Ideally, “Pericyte/SMC” should express “ACTA2” at the end of culture.