Conversion of human fibroblasts to angioblast-like progenitor cells
DOI: 10.1038/NMETH.2255
Authors: Leo Kurian; Ignacio Sancho-Martinez; Emmanuel Nivet; Aitor Aguirre; Krystal Moon; Caroline Pendaries; Cecile Volle-Challier; Francoise Bono; Jean-Marc Herbert; Julian Pulecio; Yun Xia; Mo Li; Nuria Montserrat; Sergio Ruiz; Ilir Dubova; Concepcion Rodriguez; Ahmet M Denli; Francesca S Boscolo; Rathi D Thiagarajan; Fred H Gage; Jeanne F Loring; Louise C Laurent; Juan Carlos Izpisua Belmonte
Step 1: iPSC/ESC maintainance
day 0 ~ 1
culture “adherent” human PSC in “mTeSR” for “1” days. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should be changed once in “1” days.
Ideally, “PSC” should be obtained at the end of culture.
Step 2: mesoderm 1
day 1 ~ 3
culture “adherent” human PSC in “DMEM/F12” for “2” days. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “2” days.
| concentration | unit | |
|---|---|---|
| BSA | 15 | g/l |
| insulin | 0.0175 | g/l |
| Transferrin | 0.275 | g/l |
| FGF | 2.0e-5 | g/l |
| VEGF | 5.0e-5 | g/l |
| BMP4 | 2.5e-5 | g/l |
| MTG | 4.5e-4 | mol/l |
| Glutamine | 0.00225 | mol/l |
| NEAA | 0.00225 | mol/l |
Ideally, “mesoderm” should be obtained at the end of culture.
Step 3: endothelial 1
day 3 ~ 4
culture “adherent” human mesoderm in “EBM-2” for “1” 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 “1” days.
| concentration | unit | |
|---|---|---|
| EGM-2 SingleQuots | — | — |
Ideally, “EC” should be obtained at the end of culture.