Three-Dimensional Human iPSC-Derived Artificial Skeletal Muscles Model Muscular Dystrophies and Enable Multilineage Tissue Engineering
DOI: 10.1016/j.celrep.2018.03.091
Authors: Sara Martina Maffioletti; Shilpita Sarcar; Alexander B.H. Henderson; Ingra Mannhardt; Luca Pinton; Louise Anne Moyle; Heather Steele-Stallard; Ornella Cappellari; Kim E. Wells; Giulia Ferrari; Jamie S. Mitchell; Giulia E. Tyzack; Vassilios N. Kotiadis; Moustafa Khedr; Martina Ragazzi; Weixin Wang; Michael R. Duchen; Rickie Patani; Peter S. Zammit; Dominic J. Wells; Thomas Eschenhagen; Francesco Saverio Tedesco
Step 1: endothelial 1
day 0 ~ 2
culture “device” human EC in “EC-SFM” for “2” days. For ECM, use “fibrin” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “2” days.
| concentration | unit | |
|---|---|---|
| aprotinin | 33 | g/l |
| platelet-poor plasma-derived serum | 1 | % |
| VEGF | 3.0e-5 | g/l |
| FGF | 2.0e-5 | g/l |
| HIDEM proliferation medium | 50 | % |
Ideally, “EC” should express “PECAM1” at the end of culture.
Step 2: endothelial 2
day 2 ~ 4
culture “device” human EC in “Lonza CC-3162” for “2” days. For ECM, use “fibrin” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “2” days.
| concentration | unit | |
|---|---|---|
| aprotinin | 33 | g/l |
| HIDEM differentiation medium | 50 | % |
Ideally, “EC” should express “PECAM1” at the end of culture.