The need for in vitro models that mimic the human brain to replace animal testing and allow high-throughput screening has driven scientists to develop new tools that reproduce tissue-like features on a chip. Three-dimensional (3D) in vitro cultures are emerging as an unmatched platform that preserves the complexity of cell-to-cell connections within a tissue, improves cell survival, and boosts neuronal differentiation. In this context, new and flexible imaging approaches are required to monitor the functional states of 3D networks. Herein, we propose an experimental model based on 3D neuronal networks in an alginate hydrogel, a tunable wide-volume imaging approach, and an efficient denoising algorithm to resolve, down to single cell resolution, the 3D activity of hundreds of neurons expressing the calcium sensor GCaMP6s. Furthermore, we implemented a 3D co-culture system mimicking the contiguous interfaces of distinct brain tissues such as the cortical-hippocampal interface. The analysis of the network activity of single and layered neuronal co-cultures revealed cell-type-specific activities and an organization of neuronal subpopulations that changed in the two culture configurations. Overall, our experimental platform represents a simple, powerful and cost-effective platform for developing and monitoring living 3D layered brain tissue on chip structures with high resolution and high throughput.

Fast wide-volume functional imaging of engineered in vitro brain tissues / G. Palazzolo, M. Moroni, A. Soloperto, G. Aletti, G. Naldi, M. Vassalli, T. Nieus, F. Difato. - In: SCIENTIFIC REPORTS. - ISSN 2045-2322. - 7:1(2017), pp. 8499.1-8499.20. [10.1038/s41598-017-08979-8]

Fast wide-volume functional imaging of engineered in vitro brain tissues

G. Aletti;G. Naldi;T. Nieus
Penultimo
;
2017

Abstract

The need for in vitro models that mimic the human brain to replace animal testing and allow high-throughput screening has driven scientists to develop new tools that reproduce tissue-like features on a chip. Three-dimensional (3D) in vitro cultures are emerging as an unmatched platform that preserves the complexity of cell-to-cell connections within a tissue, improves cell survival, and boosts neuronal differentiation. In this context, new and flexible imaging approaches are required to monitor the functional states of 3D networks. Herein, we propose an experimental model based on 3D neuronal networks in an alginate hydrogel, a tunable wide-volume imaging approach, and an efficient denoising algorithm to resolve, down to single cell resolution, the 3D activity of hundreds of neurons expressing the calcium sensor GCaMP6s. Furthermore, we implemented a 3D co-culture system mimicking the contiguous interfaces of distinct brain tissues such as the cortical-hippocampal interface. The analysis of the network activity of single and layered neuronal co-cultures revealed cell-type-specific activities and an organization of neuronal subpopulations that changed in the two culture configurations. Overall, our experimental platform represents a simple, powerful and cost-effective platform for developing and monitoring living 3D layered brain tissue on chip structures with high resolution and high throughput.
Computational biophysics; Imaging and sensing; Neural circuits; Tissue engineering
Settore BIO/09 - Fisiologia
Settore FIS/07 - Fisica Applicata(Beni Culturali, Ambientali, Biol.e Medicin)
Settore MAT/08 - Analisi Numerica
Settore MAT/06 - Probabilita' e Statistica Matematica
Settore SECS-S/01 - Statistica
Centro di Ricerca Interdisciplinare su Modellistica Matematica, Analisi Statistica e Simulazione Computazionale per la Innovazione Scientifica e Tecnologica ADAMSS
Article (author)
File in questo prodotto:
File Dimensione Formato  
s41598-017-08979-8.pdf

accesso aperto

Tipologia: Publisher's version/PDF
Dimensione 7.07 MB
Formato Adobe PDF
7.07 MB Adobe PDF Visualizza/Apri
Pubblicazioni consigliate

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/521276
Citazioni
  • ???jsp.display-item.citation.pmc??? 12
  • Scopus 20
  • ???jsp.display-item.citation.isi??? 22
social impact