Engraftment and morphological development of vascularized human iPS cell-derived 3D-cardiomyocyte tissue after xenotransplantation
DOI: 10.1038/s41598-017-14053-0
Authors: Hirokazu Narita; Fumiaki Shima; Junya Yokoyama; Shigeru Miyagawa; Yoshinari Tsukamoto; Yasushi Takamura; Ayami Hiura; Ken Fukumoto; Tomohiro Chiba; Seiji Watanabe; Yoshiki Sawa; Mitsuru Akashi; Hiroshi Shimoda
Step 1: iPSC/ESC maintainance
day 0 ~ 1
culture “device” human PSC in “E8” for “1” days. Medium should contain factors shown in the following table, and should be changed once in “1” days.
| concentration | unit | |
|---|---|---|
| Y27632 | 1.0e-5 | mol/l |
Ideally, “PSC” should be obtained at the end of culture.
Step 2: Primitive streak 1
day 1 ~ 2
culture “spheroid” human PSC in “StemPro” for “1” days. Medium should contain factors shown in the following table, and should be changed once in “1” days.
| concentration | unit | |
|---|---|---|
| Ascorbic Acid | 0.05 | g/l |
| Glutamine | 0.002 | mol/l |
| MTG | 4.0e-4 | mol/l |
| BMP4 | 5.0e-7 | g/l |
Step 3: mesoderm 1
day 2 ~ 7
culture “spheroid” human PSC in “StemPro” for “5” days. Medium should contain factors shown in the following table, and should be changed once in “5” days.
| concentration | unit | |
|---|---|---|
| Ascorbic Acid | 0.05 | g/l |
| Glutamine | 0.002 | mol/l |
| MTG | 4.0e-4 | mol/l |
| BMP4 | 1.0e-5 | g/l |
| FGF | 5.0e-6 | g/l |
| Activin A | 3.0e-6 | g/l |
Step 4: mesoderm 2
day 7 ~ 9
culture “spheroid” human PSC in “StemPro” for “2” days. Medium should contain factors shown in the following table, and should be changed once in “2” days.
| concentration | unit | |
|---|---|---|
| Ascorbic Acid | 0.05 | g/l |
| Glutamine | 0.002 | mol/l |
| MTG | 4.0e-4 | mol/l |
| IWR1 | 4.0e-6 | mol/l |