Blocking autocrine VEGF signaling by sunitinib, an anti-cancer drug, promotes embryonic stem cell self-renewal and somatic cell reprogramming
DOI: 10.1038/cr.2014.112
Authors: Guofang Chen; Xinxiu Xu; Lihong Zhang; Yanbin Fu; Min Wang; Haifeng Gu; Xin Xie
Step 1: iPSC/ESC maintainance
day 0 ~ 1
culture “adherent” mouse PSC in “mES medium” for “1” days. For ECM, use “Gelatin” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “1” days.
| concentration | unit | |
|---|---|---|
| LIF | 1.0e+6 | U/l |
| PD0325901 | 3.0e-6 | mol/l |
| CHIR90021 | 3.0e-6 | mol/l |
| sunitinib | 1.0e-6 | mol/l |
| DC101 | 0.02 | g/l |
| FBS | 15 | % |
| GlutaMAX | 0.002 | mol/l |
| NEAA | 1.0e-4 | mol/l |
| 2-mercaptoethanol | 1.0e-4 | mol/l |
| PSA | 100000 | U/l |
| streptomycin | 0.1 | g/l |
Ideally, “PSC” should be obtained at the end of culture.
Step 2: endothelial 1
day 1 ~ 2 (duration missing in database; assumed 1 day)
culture “adherent” mouse ESC in “unspecified medium”. For ECM, use “Gelatin” at “N/A” “N/A”. Medium should contain factors shown in the following table.
| concentration | unit | |
|---|---|---|
| VEGF | 5.0e-5 | g/l |
Ideally, “EC” should be obtained at the end of culture.