Directing stem cell fate using tuneable biomimetic peptide hydrogels

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Tissue stiffness plays a crucial role in controlling cell function and, in particular, directing stem cell fate, with seminal studies demonstrating that by matching substrate stiffness to tissue stiffness, adult human mesenchymal stem cell (MSC) differentiation can be directed down discrete lineages e.g. osteogenesis on stiff substrates vs. neurogenesis on soft substrates. This material-induced lineage commitment offers huge potential both for understanding mechanosensing/mechanotransduction mechanisms and for cell-based tissue engineering and regenerative medicine therapies. However, commonly used 2D substrates do not offer a true representation of the 3D in vivo microenvironment. Therefore this project will employ self-assembling peptide-based mechanically-tuneable biomaterials, which offer a more biomimetic environment in which to study the impact of stiffness on MSC fate. Hydrogels will be designed, synthesised and characterised in collaboration with Biogelx, a company specialising in development of chemically and mechanically tuneable self-assembling peptide hydrogels with similar nanoscale structures to human tissues, which have a wide range of applications in cell culture/biology and cell-based regenerative therapies.The project will focus on elucidating how hydrogel stiffness can be used to direct MSC lineage commitment towards soft and hard tissue phenotypes. The data generated will enhance understanding of the biological principles underpinning tissue stiffness effects on MSC fate and enable development of future regenerative therapies using hydrogels.The project will offer state-of-the-art multidisciplinary training, including biomaterial characterisation methods (atomic force microscopy, rheology, SEM and mass spectrometry); MSC phenotyping (quantitative PCR, multi-parametric flow cytometry, western blotting, immunofluorescence); transcriptomics (RNA-Seq); cell transformation and real-time confocal live-cell imaging.Richardson SM, Hughes N, Hunt JA, Freemont AJ, Hoyland JA. Human mesenchymal stem cell differentiation to NP-like cells in chitosan-glycerophosphate hydrogels. Biomaterials. 2008;29(1):85-93. Jayawarna V, Richardson SM, Hirst AR, Hodson NW, Saiani A, Gough JE, Ulijn RV. Introducing chemical functionality in Fmoc-peptide gels for cell culture. Acta Biomaterialia. 2009;5(3):934-43.Engler AJ, Sen S, Sweeney HL, Discher D.E. Matrix elasticity directs stem cell lineage specification. Cell. 2006;126(4):677-89.Swift, J., Ivanovska, I., Buxboim, A., Harada, T., Dingal, P., Pinter, J., Pajerowski, J., Spinler, K., Shin, J., Tewari, M. & Discher, D.E. Nuclear lamin-A scales with tissue stiffness and enhances matrix-directed differentiation. Science. 2013;341(6149):1240104