Direct Reprogramming of Murine Fibroblasts to Hematopoietic Progenitor Cells
DOI: 10.1016/j.celrep.2014.11.002
Authors: Kiran Batta; Magdalena Florkowska; Valerie Kouskoff; Georges Lacaud
Step 1: endothelial 1
day 0 ~ 5
culture “adherent” mouse Fibroblast in “IMDM” for “5” 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 “5” days.
| concentration | unit | |
|---|---|---|
| PDS | — | — |
| PFM | 10 | % |
| Ascorbic Acid | 5.0e-4 | mol/l |
| MTG | 4.5e-4 | mol/l |
| Glutamine | 0.002 | mol/l |
| Transferrin | 80 | g/l |
| SCF | 1 | % |
| IL-3 | 1 | % |
| GM-CSF | 1 | % |
| TPO | 1 | % |
| EPO | 4000 | U/l |
| M-CSF | 1.0e-5 | g/l |
| IL-6 | 1.0e-5 | g/l |
| IL-11 | 5.0e-6 | g/l |
| PSA | 0.05 | g/l |
| diethylaminoethyl dextran | 0.01 | g/l |
Ideally, “EC” should express “CDH5”, “TEK”, “Icam1”, “PECAM1”, “Vwf” at the end of culture.