Changes of plasmalogen phospholipid levels during differentiation of induced pluripotent stem cells 409B2 to endothelial phenotype cells
DOI: 10.1038/s41598-017-09980-x
Authors: Yusuke Nakamura; Yasuo Shimizu; Yasuhiro Horibata; Rinna Tei; Ryosuke Koike; Meitetsu Masawa; Taiji Watanabe; Taichi Shiobara; Ryo Arai; Kazuyuki Chibana; Akihiro Takemasa; Hiroyuki Sugimoto; Yoshiki Ishii
Step 1: iPSC/ESC maintainance
day 0 ~ 2
culture “adherent” human PSC in “StemFit medium” for “2” days. For ECM, use “laminin-511 E8” at “N/A” “N/A”. Medium should be changed once in “2” days.
Ideally, “PSC” should express “SSEA-4”, “alkaline phosphatase” at the end of culture.
Step 2: mesoderm 1
day 2 ~ 3 (duration missing in database; assumed 1 day)
culture “unspecified method” unknown species cell in “unspecified medium”.
Ideally, “mesoderm” should express “KDR” at the end of culture.
Step 3: endothelial 1
day 3 ~ 5
culture “adherent” human PSC in “Advanced RPMI 1640” for “2” days. For ECM, use “laminin-511 E8” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “2” days.
| concentration | unit | |
|---|---|---|
| FGF | 1.0e-5 | g/l |
| VEGF | 5.0e-5 | g/l |
| BMP4 | 2.0e-6 | g/l |
| Matrigel | 1 | % |
| B-27 Minus Insulin | 2 | % |
| GlutaMAX | 0.2 | mol/l |
Ideally, “EC” should express “KDR”, “CDH5”, “PECAM1”, “VWF” at the end of culture.
Step 4: endothelial 2
day 5 ~ 6 (duration missing in database; assumed 1 day)
culture “adherent” human iEC in “VEGF Comp Kit medium”. For ECM, use “collagen type I” at “N/A” “N/A”.
Ideally, “EC” should express “KDR”, “CDH5”, “PECAM1”, “VWF” at the end of culture.