Manipulating and functionalising personalised hydrogels to promote spinal stem cell migration and differentiation
Lead Research Organisation:
University of Leeds
Department Name: Mechanical Engineering
Abstract
Various 3D models of the spinal cord exist, including an advanced tissue engineering construct utilising collagen as matrix. These 3D models are currently being used to model spinal cord cell responses to metal wear particles from total disc replacements in the spine, and to the mechanical forces associated with spinal cord movement and injury. The responses of cells in the collagen model are readily monitored and the environment can be controlled to some extent, but there are limitations with using collagen as a matrix. We propose to develop an alternative 3D advanced hydrogel matrix that can be used as a homogenous, chemically defined and sustainable alternative to collagen gels in the construction of CNS models. A specific goal is to be able to develop a personalised gel, derived from the recipient's own tissues1, and tune the matrix physical and chemical properties in order to investigate and subsequently direct the differentiation of stem cells into neural cells. It is envisioned that by understanding the effect of personalised matrix stiffness and functionalisation on stem cell differentiation this project will lead to advances in neural tissue repair and regeneration.
Topic Area Spinal cord regeneration and repair
Outline
We will synthesise different hydrogels from donor tissues1, which will be decellularised and processed to generate a thermoresponsive hydrogel. These gels will be functionalised with biological molecules to aid cell adhesion e.g. amines and peptides. Spinal cord stem cells, which are ependymal cells (EC), will be isolated from rodent spinal cord, seeded into hydrogels and the stiffness of the hydrogels varied to determine the effect of stiffness on EC differentiation. Compounds influencing stem cell production and differentiation will be added to the cultures.
Experimental methodologies likely to be utilised during the project include cell culture, immunofluorescence, immunocytochemistry, cell viability assays, electron and confocal microscopy, 3D image analysis, ELISA, polymer synthesis, rheology and nano-indentation using AFM .
Objectives
The aims of this project are to investigate the use of different personalised hydrogels, with defined matrix stiffness and functionalised with biological additives in order to enhance cell attachment, in stem cell differentiation to create a 3D spinal cord cell model. Gel constructs will be used to investigate the effect of matrix stiffness on stem cell differentiation, neural cell phenotype and neuron-glial interactions.
Topic Area Spinal cord regeneration and repair
Outline
We will synthesise different hydrogels from donor tissues1, which will be decellularised and processed to generate a thermoresponsive hydrogel. These gels will be functionalised with biological molecules to aid cell adhesion e.g. amines and peptides. Spinal cord stem cells, which are ependymal cells (EC), will be isolated from rodent spinal cord, seeded into hydrogels and the stiffness of the hydrogels varied to determine the effect of stiffness on EC differentiation. Compounds influencing stem cell production and differentiation will be added to the cultures.
Experimental methodologies likely to be utilised during the project include cell culture, immunofluorescence, immunocytochemistry, cell viability assays, electron and confocal microscopy, 3D image analysis, ELISA, polymer synthesis, rheology and nano-indentation using AFM .
Objectives
The aims of this project are to investigate the use of different personalised hydrogels, with defined matrix stiffness and functionalised with biological additives in order to enhance cell attachment, in stem cell differentiation to create a 3D spinal cord cell model. Gel constructs will be used to investigate the effect of matrix stiffness on stem cell differentiation, neural cell phenotype and neuron-glial interactions.
Planned Impact
Regenerative Medicine been defined as "an interdisciplinary approach, spanning tissue
engineering, stem cell biology, gene therapy, cellular therapeutics, biomaterials (scaffolds and matrices),nanoscience, bioengineering and chemical biology that seeks to repair or replace damaged or diseased human cells or tissues to restore normal function, (UK Strategy for Regenerative Medicine). CDT TERM will focus on acellular therapies, scaffolds,autologous cells and regenerative devices, which can be delivered to patients as class three device interventions, thus reducing the time and cost of translation and which provide an opportunity to deliver economic growth and benefits to health in the next decade. The primary beneficiaries of CDT TERM are patients, the health service, UK industry, as well as the academic community and the students themselves. Recognising that the impact and benefit from CDT TERM will arise in the future, the statements describing impact below are supported by evidence of actual impact from our existing research and training.
Patients will benefit from regenerative interventions, which address unmet clinical needs, have improved safety and reliability, have been stratified to meet patients needs and manufactured in a cost effective manner. An example of impact arising from previous students work is a new acellular scaffold for young adult heart valve repair, which has demonstrated improved clinical outcomes at five years.
The Health Service will benefit from collaborations on research, development and evaluation of technologies, through existing partnerships with National Health Service Blood and Transplant NHSBT and the Leeds Biomedical Musculoskeletal Research Unit LMBRU. NHSBT will benefit through collaborative projects, through technology transfer, through enhancement of manufacturing processes, through pre-clinical evaluation of products and supply of trained personnel. We currently collaborate on heart valves, skin, ligaments and arteries, have licensed patents on acellular bioprocesses, and support product and process developments with pre-clinical testing and simulation. LMBRU and NHS clinicians will benefits from our collaborative research and training environment and access to our research expertise, facilities and students. Existing collaborative projects include, delivery devices for minimally manipulated stem cells and applied imaging for early OA.
Industry will benefit from supply of highly trained multidisciplinary engineers and scientists, from collaborative research and development projects, from creation and translation of IP, creation of spinout companies and through access to unique equipment, facilities and expertise. We have demonstrated: successful spin outs in form of Tissue Regenix and Credentis; successful commercialisation of a novel biological scaffolds for vascular patch repair; sustainable long term R and D and successful licensing of technology with DePuy; collaborative research with Invibio, partnering with Simulation Solutions to develop new pre-clinical simulation systems, which been adopted by regulatory agencies such as China FDA. Our graduates and researchers are employed by our industry partners.
The academic community will benefit through collaborative research and access to our facilities. We have funded collaborations with over 30 academic institutions in UK and internationally. The CDT TERM will support these collaborations and the academic partners will support student research and training. The CDT students will benefit from enhanced integrated multidisciplinary training and research, a cohort experience focused on research innovation and translation, access to our research partners, industry and clinicians. Feedback from existing students has identified the benefit of the multidisciplinary experience, the depth and breadth of excellence in our research base, the outstanding facilities and the added value of the cohort training.
engineering, stem cell biology, gene therapy, cellular therapeutics, biomaterials (scaffolds and matrices),nanoscience, bioengineering and chemical biology that seeks to repair or replace damaged or diseased human cells or tissues to restore normal function, (UK Strategy for Regenerative Medicine). CDT TERM will focus on acellular therapies, scaffolds,autologous cells and regenerative devices, which can be delivered to patients as class three device interventions, thus reducing the time and cost of translation and which provide an opportunity to deliver economic growth and benefits to health in the next decade. The primary beneficiaries of CDT TERM are patients, the health service, UK industry, as well as the academic community and the students themselves. Recognising that the impact and benefit from CDT TERM will arise in the future, the statements describing impact below are supported by evidence of actual impact from our existing research and training.
Patients will benefit from regenerative interventions, which address unmet clinical needs, have improved safety and reliability, have been stratified to meet patients needs and manufactured in a cost effective manner. An example of impact arising from previous students work is a new acellular scaffold for young adult heart valve repair, which has demonstrated improved clinical outcomes at five years.
The Health Service will benefit from collaborations on research, development and evaluation of technologies, through existing partnerships with National Health Service Blood and Transplant NHSBT and the Leeds Biomedical Musculoskeletal Research Unit LMBRU. NHSBT will benefit through collaborative projects, through technology transfer, through enhancement of manufacturing processes, through pre-clinical evaluation of products and supply of trained personnel. We currently collaborate on heart valves, skin, ligaments and arteries, have licensed patents on acellular bioprocesses, and support product and process developments with pre-clinical testing and simulation. LMBRU and NHS clinicians will benefits from our collaborative research and training environment and access to our research expertise, facilities and students. Existing collaborative projects include, delivery devices for minimally manipulated stem cells and applied imaging for early OA.
Industry will benefit from supply of highly trained multidisciplinary engineers and scientists, from collaborative research and development projects, from creation and translation of IP, creation of spinout companies and through access to unique equipment, facilities and expertise. We have demonstrated: successful spin outs in form of Tissue Regenix and Credentis; successful commercialisation of a novel biological scaffolds for vascular patch repair; sustainable long term R and D and successful licensing of technology with DePuy; collaborative research with Invibio, partnering with Simulation Solutions to develop new pre-clinical simulation systems, which been adopted by regulatory agencies such as China FDA. Our graduates and researchers are employed by our industry partners.
The academic community will benefit through collaborative research and access to our facilities. We have funded collaborations with over 30 academic institutions in UK and internationally. The CDT TERM will support these collaborations and the academic partners will support student research and training. The CDT students will benefit from enhanced integrated multidisciplinary training and research, a cohort experience focused on research innovation and translation, access to our research partners, industry and clinicians. Feedback from existing students has identified the benefit of the multidisciplinary experience, the depth and breadth of excellence in our research base, the outstanding facilities and the added value of the cohort training.
