|
|
Blood, Vol. 111, Issue 11, 5298-5306, June 1, 2008
Generation of functional platelets from human embryonic stem cells in vitro via ES-sacs, VEGF-promoted structures that concentrate hematopoietic progenitors
Blood Takayama et al.
111: 5298
Supplemental materials for: Takayama et al
Files in this Data Supplement:
- Figure S1. Hematopoietic colony formation from ES-sacs (JPG, 31.1 KB)
-
Cells obtained from ES-sacs on day 15 were cultured in Metho-Cult GF H4434 (Stem Cell Technologies, Vancouver, Canada) for an additional 14 days at 37°C. Colonies were then counted, and the counts were confirmed by Hemacolor staining. Panel A shows a representative mixed colony. Panel B summarizes the numbers of colonies arising from 1 × 104 hematopoietic cells within ES-sacs. n, neutrophils; m, macrophages; nm, neutrophils and macrophages; E, erythrocytes; Mix, mixed lineages.
- Figure S2. VEGF promotes c-Mpl expression on hematopoietic progenitors within ES-sacs (JPG, 23.9 KB)
-
The hematopoietic progenitors within ES-sacs on day 15 were collected and stained with anti-human c-Mpl antibody or control mouse IgG, following by staining with Alexa 488-conjugated anti-mouse IgG secondary antibody and flow cytometric analysis. Numbers of cells showing c-Mpl expression in the absence and presence of 20 ng/ml VEGF were calculated using True Count Beads.
- Video 1. Time-lapse video of ES-sac formation (MOV, 6.13 MB)
-
Human ESCs were seeded onto C3H10T1/2 cells on day 0. Time-lapse images were taken through a phase-contrast microscope inside a culture system at 37°C under a 5% CO2 /95% air atmosphere (MCOK-5M; Sanyo, Tokyo, Japan). Time-lapse recording was started on day 4 and continued until day 12 in the presence of 20 ng/ml VEGF. Recorded images were accumulated on a computer using MTR-4000 software (Sanyo).
|
|
