Production of 3-dimensional neural progenitor cultures from human pluripotent stem
cells offers the potential to generate large numbers of cells. We utilised our nanobridge
system to generate 3D hPSC aggregates for differentiation towards the neural lineage,
and investigate the ability to passage aggregates while maintaining cells at a stem/progenitor
stage. Over 38 days, aggregate cultures exhibited upregulation and maintenance of
neural-associated markers and demonstrated up to 10 fold increase in cell number.
Aggregates undergoing neural induction in the presence or absence of nanobridges demonstrated
no differences in marker expression, proliferation or viability. However, aggregates
formed without nanobridges were statistically significantly fewer and smaller by passage
3. Organoids, cultured from aggregates, and treated with retinoic acid or rock inhibitor
demonstrated terminal differentiation as assessed by immunohistochemistry. These data
demonstrate that nanobridge 3D hPSC can differentiate to neural stem/progenitor cells,
and be maintained at this stage through serial passaging and expansion.