Human dental pulp pluripotent-like stem cells promote wound healing and muscle regeneration
DOI: 10.1186/s13287-017-0621-3
Authors: Ester Martínez-Sarrà; Sheyla Montori; Carlos Gil-Recio; Raquel Núñez-Toldrà; Domiziana Costamagna; Alessio Rotini; Maher Atari; Aernout Luttun; Maurilio Sampaolesi
Step 1: iPSC/ESC maintainance
day 0 ~ 4
culture “adherent” human DPPSC in “DMEM-low glucose” for “4” days. For ECM, use “FN1” at “1.0e-4” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “4” days.
| concentration | unit | |
|---|---|---|
| MCDB-201 | 40 | % |
| ITS | 1 | x |
| LA-BSA | 1 | x |
| Dexamethasone | 1.0e-9 | mol/l |
| Ascorbic Acid | 1.0e-4 | mol/l |
| PSA | 100 | U |
| streptomycin | 1000 | U |
| FBS | 2 | % |
| PDGF-BB | 1.0e-5 | g/l |
| EGF | 1.0e-5 | g/l |
| LIF | 1.0e+6 | U/l |
| Chemically Defined Lipid Concentrate | — | — |
| BSA | 0.8 | g/l |
| 2-mercaptoethanol | 0.055 | mol/l |
Ideally, “DPPSC” should express “OCT4A”, “NANOG”, “SOX2”, “ALP” at the end of culture.
Step 2: endothelial 1
day 4 ~ 6
culture “adherent” human DPPSC in “EGM2” for “2” days. Medium should be changed once in “2” days.
Ideally, “EC” should express “VWF”, “PECAM1”, “CDH5”, “KDR” at the end of culture.