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Customizable, engineered substrates for rapid screening of cellular cues
Biophysical cues robustly direct cell responses and are thus important tools for in vitro and translational biomedical applications. High throughput platforms exploring substrates with varying physical properties are therefore valuable. However, currently existing platforms are limited in throughput...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
IOP Publishing
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7655147/ https://www.ncbi.nlm.nih.gov/pubmed/31783378 http://dx.doi.org/10.1088/1758-5090/ab5d3f |
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author | Huethorst, Eline Cutiongco, Marie FA Campbell, Fraser A Saeed, Anwer Love, Rachel Reynolds, Paul M Dalby, Matthew J Gadegaard, Nikolaj |
author_facet | Huethorst, Eline Cutiongco, Marie FA Campbell, Fraser A Saeed, Anwer Love, Rachel Reynolds, Paul M Dalby, Matthew J Gadegaard, Nikolaj |
author_sort | Huethorst, Eline |
collection | PubMed |
description | Biophysical cues robustly direct cell responses and are thus important tools for in vitro and translational biomedical applications. High throughput platforms exploring substrates with varying physical properties are therefore valuable. However, currently existing platforms are limited in throughput, the biomaterials used, the capability to segregate between different cues and the assessment of dynamic responses. Here we present a multiwell array (3 × 8) made of a substrate engineered to present topography or rigidity cues welded to a bottomless plate with a 96-well format. Both the patterns on the engineered substrate and the well plate format can be easily customized, permitting systematic and efficient screening of biophysical cues. To demonstrate the broad range of possible biophysical cues examinable, we designed and tested three multiwell arrays to influence cardiomyocyte, chondrocyte and osteoblast function. Using the multiwell array, we were able to measure different cell functionalities using analytical modalities such as live microscopy, qPCR and immunofluorescence. We observed that grooves (5 μm in size) induced less variation in contractile function of cardiomyocytes. Compared to unpatterned plastic, nanopillars with 127 nm height, 100 nm diameter and 300 nm pitch enhanced matrix deposition, chondrogenic gene expression and chondrogenic maintenance. High aspect ratio pillars with an elastic shear modulus of 16 kPa mimicking the matrix found in early stages of bone development improved osteogenic gene expression compared to stiff plastic. We envisage that our bespoke multiwell array will accelerate the discovery of relevant biophysical cues through improved throughput and variety. |
format | Online Article Text |
id | pubmed-7655147 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | IOP Publishing |
record_format | MEDLINE/PubMed |
spelling | pubmed-76551472020-11-12 Customizable, engineered substrates for rapid screening of cellular cues Huethorst, Eline Cutiongco, Marie FA Campbell, Fraser A Saeed, Anwer Love, Rachel Reynolds, Paul M Dalby, Matthew J Gadegaard, Nikolaj Biofabrication Paper Biophysical cues robustly direct cell responses and are thus important tools for in vitro and translational biomedical applications. High throughput platforms exploring substrates with varying physical properties are therefore valuable. However, currently existing platforms are limited in throughput, the biomaterials used, the capability to segregate between different cues and the assessment of dynamic responses. Here we present a multiwell array (3 × 8) made of a substrate engineered to present topography or rigidity cues welded to a bottomless plate with a 96-well format. Both the patterns on the engineered substrate and the well plate format can be easily customized, permitting systematic and efficient screening of biophysical cues. To demonstrate the broad range of possible biophysical cues examinable, we designed and tested three multiwell arrays to influence cardiomyocyte, chondrocyte and osteoblast function. Using the multiwell array, we were able to measure different cell functionalities using analytical modalities such as live microscopy, qPCR and immunofluorescence. We observed that grooves (5 μm in size) induced less variation in contractile function of cardiomyocytes. Compared to unpatterned plastic, nanopillars with 127 nm height, 100 nm diameter and 300 nm pitch enhanced matrix deposition, chondrogenic gene expression and chondrogenic maintenance. High aspect ratio pillars with an elastic shear modulus of 16 kPa mimicking the matrix found in early stages of bone development improved osteogenic gene expression compared to stiff plastic. We envisage that our bespoke multiwell array will accelerate the discovery of relevant biophysical cues through improved throughput and variety. IOP Publishing 2020-04 2020-02-07 /pmc/articles/PMC7655147/ /pubmed/31783378 http://dx.doi.org/10.1088/1758-5090/ab5d3f Text en © 2020 IOP Publishing Ltd http://creativecommons.org/licenses/by/3.0/ Original content from this work may be used under the terms of the Creative Commons Attribution 3.0 licence (http://creativecommons.org/licenses/by/3.0) . Any further distribution of this work must maintain attribution to the author(s) and the title of the work, journal citation and DOI. |
spellingShingle | Paper Huethorst, Eline Cutiongco, Marie FA Campbell, Fraser A Saeed, Anwer Love, Rachel Reynolds, Paul M Dalby, Matthew J Gadegaard, Nikolaj Customizable, engineered substrates for rapid screening of cellular cues |
title | Customizable, engineered substrates for rapid screening of cellular
cues |
title_full | Customizable, engineered substrates for rapid screening of cellular
cues |
title_fullStr | Customizable, engineered substrates for rapid screening of cellular
cues |
title_full_unstemmed | Customizable, engineered substrates for rapid screening of cellular
cues |
title_short | Customizable, engineered substrates for rapid screening of cellular
cues |
title_sort | customizable, engineered substrates for rapid screening of cellular
cues |
topic | Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7655147/ https://www.ncbi.nlm.nih.gov/pubmed/31783378 http://dx.doi.org/10.1088/1758-5090/ab5d3f |
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