Customizable, engineered substrates for rapid screening of cellular cues

Huethorst, E., Cutiongco, M. F.A. , Campbell, F. A., Saeed, A. , Love, R., Reynolds, P. M. , Dalby, M. J. and Gadegaard, N. (2020) Customizable, engineered substrates for rapid screening of cellular cues. Biofabrication, 12(2), 025009. (doi: 10.1088/1758-5090/ab5d3f) (PMID:31783378)

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Abstract

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 (3x8) 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.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Saeed, Dr Anwer and Huethorst, Eline and Cutiongco, Ms Marie and Love, Miss Rachel and Dalby, Professor Matthew and Campbell, Fraser and Reynolds, Dr Paul and Gadegaard, Professor Nikolaj
Authors: Huethorst, E., Cutiongco, M. F.A., Campbell, F. A., Saeed, A., Love, R., Reynolds, P. M., Dalby, M. J., and Gadegaard, N.
College/School:College of Medical Veterinary and Life Sciences > Institute of Molecular Cell and Systems Biology
College of Science and Engineering > School of Engineering > Biomedical Engineering
College of Science and Engineering > School of Engineering > James Watt Nanofabrication Centre
Journal Name:Biofabrication
Publisher:IOP Publishing
ISSN:1758-5082
ISSN (Online):1758-5090
Published Online:29 November 2019
Copyright Holders:Copyright © 2019 IOP Publishing Ltd
First Published:First published in Biofabrication 12(2):025009
Publisher Policy:Reproduced under a Creative Commons License

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
190790EPSRC DTG 2014Mary Beth KneafseyEngineering and Physical Sciences Research Council (EPSRC)EP/M506539/1Research and Innovation Services
172865EPSRC DTP 16/17 and 17/18Tania GalabovaEngineering and Physical Sciences Research Council (EPSRC)EP/N509668/1Research and Innovation Services
173470BHF 4-Year PhD Studentship (4th intake 2016 of 3rd Scheme)Rhian TouyzBritish Heart Foundation (BHF)FS/16/55/32731Institute of Cardiovascular & Medical Sciences