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Stem Cell Res. 2019 Jul;38:101441. doi: 10.1016/j.scr.2019.101441. Epub 2019 Apr 23.

Fibronectin-conjugated thermoresponsive nanobridges generate three dimensional human pluripotent stem cell cultures for differentiation towards the neural lineages.

Author information

1
Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, Brisbane, QLD 4072, Australia. Electronic address: l.harkness@uq.edu.au.
2
Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, Brisbane, QLD 4072, Australia.
3
Translational Cell Imaging Queensland, Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, QLD 4102, Australia.
4
The Florey Institute of Neuroscience and Mental Health and the Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3052, Australia; The Jackson Laboratory, Bar Harbor, ME 04609, United States; The University of Melbourne, Melbourne, Victoria 3010, Australia.

Abstract

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.

KEYWORDS:

3D cell culture; Neural progenitors; Thermoresponsive polymer; hESC

PMID:
31082678
DOI:
10.1016/j.scr.2019.101441
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