Therapeutic angiogenesis by transplantation of induced pluripotent stem cell-derived Flk-1 positive cells
Authors: Hirohiko Suzuki; Rei Shibata; Tetsutaro Kito; Masakazu Ishii; Ping Li; Toru Yoshikai; Naomi Nishio; Sachiko Ito; Yasushi Numaguchi; Jun K Yamashita; Toyoaki Murohara; Kenichi Isobe
Step 1: iPSC/ESC maintainance
day 0 ~ 1 (duration missing in database; assumed 1 day)
culture “adherent” mouse PSC in “DMEM”. For ECM, use “MEF feeder” at “N/A” “N/A”. Medium should contain factors shown in the following table.
| concentration | unit | |
|---|---|---|
| KSR | 10 | % |
| FBS | 1 | % |
| NEAA | — | — |
| 2-mercaptoethanol | 0.0055 | mol/l |
| PSA | 50000 | U/l |
| streptomycin | 50 | g/l |
| LIF | — | — |
Step 2: mesoderm 1
day 1 ~ 5
culture “adherent” mouse PSC in “alpha-MEM” for “4” days. For ECM, use “Collagen” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “4” days.
| concentration | unit | |
|---|---|---|
| FBS | 10 | % |
| 2-mercaptoethanol | 5.0e-5 | mol/l |
Ideally, “mesoderm” should express “KDR”, “NANOG”, “POU5F1”, “c-myc” at the end of culture.