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

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 ag...

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Bibliographic Details
Published in:Stem cell research 2019-07, Vol.38, p.101441-101441, Article 101441
Main Authors: Harkness, Linda, Chen, Xiaoli, Jia, Zhongfan, Davies, Anthony M., Monteiro, Michael, Gray, Peter, Pera, Martin
Format: Article
Language:English
Subjects:
3D cell culture
Cell Differentiation
Cell Line
Fibronectins - chemistry
hESC
Human Embryonic Stem Cells - cytology
Human Embryonic Stem Cells - metabolism
Humans
Nanostructures - chemistry
Neural progenitors
Neural Stem Cells - cytology
Neural Stem Cells - metabolism
Thermoresponsive polymer
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Summary: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. •Fibronectin conjugated thermoresponsive polymer for production of hESC aggregates•Neural induced aggregates show high viability, proliferation, no necrotic core•Expansion of PAX6 and SOX1 positive 3D NPC during d38 neural differentiation•O4/TUBB/MAP2/GFAP staining demonstrated at 94 days of induction
ISSN:1873-5061
1876-7753
DOI:10.1016/j.scr.2019.101441