Directing stem cell fate using tuneable biomimetic peptide hydrogels
Lead Research Organisation:
University of Manchester
Department Name: School of Biological Sciences
Abstract
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
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
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| BB/M011208/1 | 01/10/2015 | 31/03/2024 | |||
| 1792803 | Studentship | BB/M011208/1 | 01/10/2016 | 31/03/2021 |
| Description | Introduction: Bone marrow mesenchymal stem cells (BMMSCs) are a population of multipotent stem cells derived from the bone marrow of long bones. BMMSCs are defined by their ability for tri-linage differentiation along adipogenic (fatty tissue), osteogenic (bone) and chondrogenic (cartilage)) lineages. Differentiation of BMMSCs is typically achieved using a cocktail of growth factors and chemicals within the cell culture medium. However, differentiation can also be achieved solely by altering the stiffness of the surface on which the BMMSCs are cultured; without the use of biological and chemical factors (Engler et al. 2006) (The field which investiagtes how cells respond to the mechanical properties of materials is known as mechanobiology). Much of the research in this field to date has focused on 2D culture. In this method BMMSCs are grown atop surfaces of different stiffness. My focus is to progress this by developing a system to investigate the effect of stiffness on BMMSCs in 3D culture. The system I am using is an Fmoc based peptide hydrogel designed by our insturial partner, Biogelx. Fmoc peptide hydrogels have been shown to support BMMSCs in 2D and 3D culture and can be created in a wide range of stiffnesses by altering the initial concentration of the pregel solution. However, they have not previously been extensively used for mechanobiology experiments. Main discoveries: 1) I have extensively characterised the mechanical properties of a range of Fmoc peptide hydrogels which have novel combinations of peptides. 2) Using molecular biological techniques, I have characterised the biological response of BMMSCs to different stiffnesses of Fmoc peptide hydrogels using 2D culture. WHile further work is required, my initial findings suggest the BMMSCs appear to respond in a similar fashion to other materials used in mechanobiology experiments. Future work will focus on translating these findings to 3D culture now I have validated my experimental system and an -Omics based pathway analysis to investigate novel mechanosensitive pathways. |
| Exploitation Route | My findings will help broaden the understanding of the response of BMMSCs to their mechanical environment. More broadly, it will help others who also use peptide hydrogels by providing further information about how BMMSCs interact with this unique type of material. |
| Sectors | Pharmaceuticals and Medical Biotechnology |
| Description | A Level Study Day |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Schools |
| Results and Impact | Voluntary public engagement activity teaching local A-Level students about stem cells at the Univerisity of Manchester Museum. |
| Year(s) Of Engagement Activity | 2018 |
| Description | RCS Biomaterials 2018 Conference Talk |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Professional Practitioners |
| Results and Impact | 5 minute talk at the RCS Biomaterials Special interest group meeting. Audience ~150 people |
| Year(s) Of Engagement Activity | 2018 |
| Description | The Body Experience (Public Engagement) |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Public/other audiences |
| Results and Impact | Voluntary public engagement activity at the University of Manchester Museum. Teaching the general public about aspects of research at the University / our lab through child-focused fun activities. |
| Year(s) Of Engagement Activity | 2017 |
| Description | UKMSC 2017 Conference Talk |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Professional Practitioners |
| Results and Impact | 10 minute talk at the UKMSC conference in 2017. Selected based on abstract. Audience ~200 people. |
| Year(s) Of Engagement Activity | 2017 |