Manipulating and functionalising personalised hydrogels to promote spinal stem cell migration and differentiation

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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 repairOutlineWe 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 .ObjectivesThe 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.