Graphene Oxide promotes embryonic stem cell differentiation to haematopoietic lineage
DOI: 10.1038/srep25917
Authors: Eva Garcia-Alegria; Maria Iliut; Monika Stefanska; Claudio Silva; Sebastian Heeg; Susan J. Kimber; Valerie Kouskoff; Georges Lacaud; Aravind Vijayaraghavan; Kiran Batta
Step 1: iPSC/ESC maintainance
day 0 ~ 1 (duration missing in database; assumed 1 day)
culture “adherent” human ESC in “E8”. For ECM, use “Geltrex” at “N/A” “N/A”.
Ideally, “ESC” should be obtained at the end of culture.
Step 2: mesoderm 2
day 1 ~ 4
culture “adherent” mouse mesoderm in “IMDM” for “3” days. For ECM, use “graphene oxide” at “1.0e-8” “mol/l”. Medium should contain factors shown in the following table, and should be changed once in “3” days.
| concentration | unit | |
|---|---|---|
| FBS | 10 | % |
| Ascorbic Acid | 5.0e-4 | mol/l |
| MTG | 4.5e-4 | mol/l |
| Glutamine | 0.002 | mol/l |
| Transferrin | 80 | g/l |
| PSA | 0.05 | g/l |
Ideally, “EC” should express “TIE2”, “VE-CADHERIN”, “CD41”, “c-KIT” at the end of culture.
Step 3: endothelial 1
day 4 ~ 6
culture “adherent” mouse EC in “IMDM” for “2” days. For ECM, use “graphene oxide” at “1.0e-8” “mol/l”. Medium should contain factors shown in the following table, and should be changed once in “2” days.
| concentration | unit | |
|---|---|---|
| plasma-derived serum | — | — |
| protein-free hybridoma medium | 10 | % |
| Ascorbic Acid | 5.0e-4 | mol/l |
| MTG | 4.5e-4 | mol/l |
| Glutamine | 0.002 | mol/l |
| Transferrin | 80 | g/l |
| PSA | 0.05 | g/l |
| IL3 | 1 | % |
| GM-CSF | 1 | % |
| M-CSF | 1.0e-5 | g/l |
| SCF | 1 | % |
| Thrombopoietin | 1 | % |
| IL11 | — | — |
| IL6 | — | — |
| Erythropoietin | 4000 | U/l |
Ideally, “EC” should express “TIE2”, “c-KIT”, “CD41” at the end of culture.