An immortalised mesenchymal stem cell line maintains mechano-responsive behaviour and can be used as a reporter of substrate stiffness

Galarza Torre, A., Shaw, J. E., Wood, A., Gilbert, H. T.J., Dobre, O. , Genever, P., Brennan, K., Richardson, S. M. and Swift, J. (2018) An immortalised mesenchymal stem cell line maintains mechano-responsive behaviour and can be used as a reporter of substrate stiffness. Scientific Reports, 8, 8981. (doi: 10.1038/s41598-018-27346-9) (PMID:29895825) (PMCID:PMC5997644)

Abstract

The mechanical environment can influence cell behaviour, including changes to transcriptional and proteomic regulation, morphology and, in the case of stem cells, commitment to lineage. However, current tools for characterizing substrates' mechanical properties, such as atomic force microscopy (AFM), often do not fully recapitulate the length and time scales over which cells 'feel' substrates. Here, we show that an immortalised, clonal line of human mesenchymal stem cells (MSCs) maintains the responsiveness to substrate mechanics observed in primary cells, and can be used as a reporter of stiffness. MSCs were cultured on soft and stiff polyacrylamide hydrogels. In both primary and immortalised MSCs, stiffer substrates promoted increased cell spreading, expression of lamin-A/C and translocation of mechano-sensitive proteins YAP1 and MKL1 to the nucleus. Stiffness was also found to regulate transcriptional markers of lineage. A GFP-YAP/RFP-H2B reporter construct was designed and virally delivered to the immortalised MSCs for in situ detection of substrate stiffness. MSCs with stable expression of the reporter showed GFP-YAP to be colocalised with nuclear RFP-H2B on stiff substrates, enabling development of a cellular reporter of substrate stiffness. This will facilitate mechanical characterisation of new materials developed for applications in tissue engineering and regenerative medicine.

• Item Type: Articles
• Additional Information: HTJG and JS were funded by a Biotechnology and Biological Sciences Research Council (BBSRC) David Phillips Fellowship (BB/L024551/1). OD was supported by a Wellcome Trust Institutional Strategic Support Fund (097820/Z/11/B). PG was funded by the National Centre for the Replacement, Refnement and Reduction of Animals in Research (NC3Rs) and Arthritis Research UK (ARUK). Microscopy was carried out at the Wellcome Centre for Cell-Matrix Research (WTCCMR; 203128/Z/16/Z) Imaging Core Facility.
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Dobre, Dr Oana
• Authors: Galarza Torre, A., Shaw, J. E., Wood, A., Gilbert, H. T.J., Dobre, O., Genever, P., Brennan, K., Richardson, S. M., and Swift, J.
• College/School: College of Science and Engineering > School of Engineering > Biomedical Engineering
• Journal Name: Scientific Reports
• Publisher: Nature Research
• ISSN: 2045-2322
• ISSN (Online): 2045-2322
• Copyright Holders: Copyright © 2018 The Authors
• First Published: First published in Scientific Reports 8: 8981
• Publisher Policy: Reproduced under a Creative Commons License