Serial passaging of hiPSCs in static suspension in E8 medium. (a) Light microscope image of aggregates after 48 h post-inoculation of single suspension BC1 cells in ultra-low attachment plates. (b) BC1 cells cultured in E8 medium, expanded in static suspension culture for 13 passages (45 days, n = 3). (c) Flow cytometry plots (n = 3) of TRA-1–60 expression in hiPSCs after 13 passages in static suspension. (d) Light microscope image of HPC-like single suspending cells emerging around a spin-EB on day 14. (e) Flow cytometry plots (n = 2) of CD34 and CD45 expression in day-14 spin EBs. NC = negative control. Scale bars = 200 μm.
aggregates
Light microscope image
after 48 h
h
BC1 cells cultured in E8 medium
expanded in static suspension culture
13 passages
passages
13 passages
45 days
13 passages
n
3
Flow cytometry plots
n
3
static suspension
after 13 passages
passages
Scale bars
200 μm
μm