Cargando…

Microfluidic Biofabrication of 3D Multicellular Spheroids by Modulation of Non-geometrical Parameters

Three-dimensional (3D) cell spheroids are being increasingly applied in many research fields due to their enhanced biological functions as compared to conventional two-dimensional (2D) cultures. 3D cell spheroids can replicate tissue functions, which enables their use both as in vitro models and as...

Descripción completa

Detalles Bibliográficos
Autores principales: Lopa, Silvia, Piraino, Francesco, Talò, Giuseppe, Mainardi, Valerio Luca, Bersini, Simone, Pierro, Margherita, Zagra, Luigi, Rasponi, Marco, Moretti, Matteo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7214796/
https://www.ncbi.nlm.nih.gov/pubmed/32432090
http://dx.doi.org/10.3389/fbioe.2020.00366
_version_ 1783532046076870656
author Lopa, Silvia
Piraino, Francesco
Talò, Giuseppe
Mainardi, Valerio Luca
Bersini, Simone
Pierro, Margherita
Zagra, Luigi
Rasponi, Marco
Moretti, Matteo
author_facet Lopa, Silvia
Piraino, Francesco
Talò, Giuseppe
Mainardi, Valerio Luca
Bersini, Simone
Pierro, Margherita
Zagra, Luigi
Rasponi, Marco
Moretti, Matteo
author_sort Lopa, Silvia
collection PubMed
description Three-dimensional (3D) cell spheroids are being increasingly applied in many research fields due to their enhanced biological functions as compared to conventional two-dimensional (2D) cultures. 3D cell spheroids can replicate tissue functions, which enables their use both as in vitro models and as building blocks in tissue biofabrication approaches. In this study, we developed a perfusable microfluidic platform suitable for robust and reproducible 3D cell spheroid formation and tissue maturation. The geometry of the device was optimized through computational fluid dynamic (CFD) simulations to improve cell trapping. Experimental data were used in turn to generate a model able to predict the number of trapped cells as a function of cell concentration, flow rate, and seeding time. We demonstrated that tuning non-geometrical parameters it is possible to control the size and shape of 3D cell spheroids generated using articular chondrocytes (ACs) as cellular model. After seeding, cells were cultured under perfusion at different flow rates (20, 100, and 500 μl/min), which induced the formation of conical and spherical spheroids. Wall shear stress values on cell spheroids, computed by CFD simulations, increased accordingly to the flow rate while remaining under the chondroprotective threshold in all configurations. The effect of flow rate on cell number, metabolic activity, and tissue-specific matrix deposition was evaluated and correlated with fluid velocity and shear stress distribution. The obtained results demonstrated that our device represents a helpful tool to generate stable 3D cell spheroids which can find application both to develop advanced in vitro models for the study of physio-pathological tissue maturation mechanisms and to obtain building blocks for the biofabrication of macrotissues.
format Online
Article
Text
id pubmed-7214796
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-72147962020-05-19 Microfluidic Biofabrication of 3D Multicellular Spheroids by Modulation of Non-geometrical Parameters Lopa, Silvia Piraino, Francesco Talò, Giuseppe Mainardi, Valerio Luca Bersini, Simone Pierro, Margherita Zagra, Luigi Rasponi, Marco Moretti, Matteo Front Bioeng Biotechnol Bioengineering and Biotechnology Three-dimensional (3D) cell spheroids are being increasingly applied in many research fields due to their enhanced biological functions as compared to conventional two-dimensional (2D) cultures. 3D cell spheroids can replicate tissue functions, which enables their use both as in vitro models and as building blocks in tissue biofabrication approaches. In this study, we developed a perfusable microfluidic platform suitable for robust and reproducible 3D cell spheroid formation and tissue maturation. The geometry of the device was optimized through computational fluid dynamic (CFD) simulations to improve cell trapping. Experimental data were used in turn to generate a model able to predict the number of trapped cells as a function of cell concentration, flow rate, and seeding time. We demonstrated that tuning non-geometrical parameters it is possible to control the size and shape of 3D cell spheroids generated using articular chondrocytes (ACs) as cellular model. After seeding, cells were cultured under perfusion at different flow rates (20, 100, and 500 μl/min), which induced the formation of conical and spherical spheroids. Wall shear stress values on cell spheroids, computed by CFD simulations, increased accordingly to the flow rate while remaining under the chondroprotective threshold in all configurations. The effect of flow rate on cell number, metabolic activity, and tissue-specific matrix deposition was evaluated and correlated with fluid velocity and shear stress distribution. The obtained results demonstrated that our device represents a helpful tool to generate stable 3D cell spheroids which can find application both to develop advanced in vitro models for the study of physio-pathological tissue maturation mechanisms and to obtain building blocks for the biofabrication of macrotissues. Frontiers Media S.A. 2020-05-05 /pmc/articles/PMC7214796/ /pubmed/32432090 http://dx.doi.org/10.3389/fbioe.2020.00366 Text en Copyright © 2020 Lopa, Piraino, Talò, Mainardi, Bersini, Pierro, Zagra, Rasponi and Moretti. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Bioengineering and Biotechnology
Lopa, Silvia
Piraino, Francesco
Talò, Giuseppe
Mainardi, Valerio Luca
Bersini, Simone
Pierro, Margherita
Zagra, Luigi
Rasponi, Marco
Moretti, Matteo
Microfluidic Biofabrication of 3D Multicellular Spheroids by Modulation of Non-geometrical Parameters
title Microfluidic Biofabrication of 3D Multicellular Spheroids by Modulation of Non-geometrical Parameters
title_full Microfluidic Biofabrication of 3D Multicellular Spheroids by Modulation of Non-geometrical Parameters
title_fullStr Microfluidic Biofabrication of 3D Multicellular Spheroids by Modulation of Non-geometrical Parameters
title_full_unstemmed Microfluidic Biofabrication of 3D Multicellular Spheroids by Modulation of Non-geometrical Parameters
title_short Microfluidic Biofabrication of 3D Multicellular Spheroids by Modulation of Non-geometrical Parameters
title_sort microfluidic biofabrication of 3d multicellular spheroids by modulation of non-geometrical parameters
topic Bioengineering and Biotechnology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7214796/
https://www.ncbi.nlm.nih.gov/pubmed/32432090
http://dx.doi.org/10.3389/fbioe.2020.00366
work_keys_str_mv AT lopasilvia microfluidicbiofabricationof3dmulticellularspheroidsbymodulationofnongeometricalparameters
AT pirainofrancesco microfluidicbiofabricationof3dmulticellularspheroidsbymodulationofnongeometricalparameters
AT talogiuseppe microfluidicbiofabricationof3dmulticellularspheroidsbymodulationofnongeometricalparameters
AT mainardivalerioluca microfluidicbiofabricationof3dmulticellularspheroidsbymodulationofnongeometricalparameters
AT bersinisimone microfluidicbiofabricationof3dmulticellularspheroidsbymodulationofnongeometricalparameters
AT pierromargherita microfluidicbiofabricationof3dmulticellularspheroidsbymodulationofnongeometricalparameters
AT zagraluigi microfluidicbiofabricationof3dmulticellularspheroidsbymodulationofnongeometricalparameters
AT rasponimarco microfluidicbiofabricationof3dmulticellularspheroidsbymodulationofnongeometricalparameters
AT morettimatteo microfluidicbiofabricationof3dmulticellularspheroidsbymodulationofnongeometricalparameters