Enhanced Lung Epithelial Specification of Human Induced Pluripotent Stem Cells on Decellularized Lung Matrix
DOI: 10.1016/j.athoracsur.2014.05.080
Authors: Sarah E. Gilpin; Xi Ren; Tatsuya Okamoto; Jacques P. Guyette; Hongmei Mou; Jayaraj Rajagopal; Douglas J. Mathisen; Joseph P. Vacanti; Harald C. Ott
Step 1: iPSC/ESC maintainance
day 0 ~ 1 (duration missing in database; assumed 1 day)
culture “adherent” human PSC in “mTeSR”. For ECM, use “Geltrex” at “N/A” “N/A”.
Ideally, “PSC” should be obtained at the end of culture.
Step 2: endothelial 1
day 1 ~ 7
culture “unspecified method” human PSC in “IMDM” for “6” days. Medium should contain factors shown in the following table, and should be changed once in “6” days.
| concentration | unit | |
|---|---|---|
| BIT 9500 | — | — |
| NEAA | — | — |
| Glutamine | — | — |
| MTG | 10 | % |
| PSA | — | — |
| BMP4 | 5.0e-5 | g/l |
| FGF | 5.0e-5 | g/l |
| VEGF | 5.0e-5 | g/l |
Ideally, “EC” should be obtained at the end of culture.
Step 3: endothelial 2
day 7 ~ 8 (duration missing in database; assumed 1 day)
culture “adherent” human EC in “EGM2”. For ECM, use “Gelatin” at “0.1” “%”. Medium should contain factors shown in the following table.
| concentration | unit | |
|---|---|---|
| FBS | 20 | % |
Ideally, “EC” should be obtained at the end of culture.