In vitro conversion of adult murine endothelial cells to hematopoietic stem cells
DOI: 10.1038/s41596-018-0060-3
Authors: José Gabriel Barcia Durán; Raphaël Lis; Tyler M. Lu; Shahin Rafii
Step 1: endothelial 1
day 0 ~ 2
culture “adherent” unknown species cell and mouse EC and human EC in “M199” for “2” days. For ECM, use “FN1” at “1” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “2” days.
| concentration | unit | |
|---|---|---|
| FBS | 10 | % |
| Heparin | 0.1 | g/l |
| endothelial mitogen | 0.05 | g/l |
| HEPES | 0.02 | mol/l |
| GlutaMAX | 1 | x |
| doxycycline | 0.001 | g/l |
| polybrene | 5.0e-6 | g |
Ideally, “EC” should express “PECAM1”, “CDH5” at the end of culture.