Culture of hiPSC-derived pancreatic cells
hiPSC-derived pancreatic cells (day 12 of differentiation) were seeded
on scaffolds with two different modifications: amine-modified and
gelatin-coated, NaOH-modified. Cells were seeded either as single cells
or as aggregates. For seeding single cells, the aforementioned method
was followed. To seed the aggregates, cell islets with an average of
4000 cells were detached manually from wells of a 6-well plate by a 10
µl pipette tip under microscopic observation and were transferred to a
15 ml centrifuge tube followed by gravitational settling. After 10
minutes at 37° C, the supernatant was removed, and aggregates were
resuspended in 360 µl of stage-four medium (DMEM/F12, B27, p/s). 10 µl
of cell suspension, containing 5 aggregates was added to each prepared
scaffold. The same differentiation protocol was followed for cells grown
on scaffolds as in dishes.