Neoteric Biomaterials for hiPSCs Monitorized Differentiation to RGCs: Creation, Microfabrication & Microfluidics (bioMAT4EYE)

I bioMAT4EYE-prosjektet skal det utvikles nye avanserte støttematerialer hvor iPSC (Induced Pluripotent StemCells) cellekultur kan vokse i, og hvor iPSC-celler kan differensieres til nye celletyper som RGC-celler (retinale ganglionceller). I den første prosjektperioden har ulike biopolymerer blitt testet som støttemateriale. RISE PFI har utviklet biopolymerblekk for 3D-trykking av scaffolds (støttestrukturer), med fokus på kitosan med varierende molekylvekt og grad av deacetylering. Scaffolds har blitt overflatemodifisert for å sikre at celler festes til scaffolds-overflaten og gjennom dette promotere at cellene differensieres til retinale ganglionceller. Foreløpige forsøk har demonstrert at de utviklede blekkene av kitosan kan 3D-trykkes, og at celler festes til de overflatemodifiserte scaffoldene av kitosan. I tillegg har Chitinor utviklet kompetanse og kommersielle produksjonsprosedyrer som muliggjør produksjon av farmasøytisk kitosan med ekstreme spesifikasjoner for molekylvekt, for best mulig tilpasning til bruk i 3D-blekk. Målet er at de nye støttematerialene skal kunne gi grunnlaget for en ny type celleterapibehandling for pasienter med skader på synsnerven (optisk nevropati).

Numerous cell culture applications have been performed on Matrigel, a basement-membrane matrix extracted from Engelbreth–Holm–Swarm (EHS) mouse sarcomas. From a chemical and structural perspective, Matrigel is of an extraordinary complexity; even if only 4 proteins and proteoglycans account for most of it, up to 15000 peptides and 2000 proteins are additional minor components. Its complexity and origin lead to a batch-to-batch scarce reproducibility, so even lab results based on such biomaterial may not be reliable. Thus, there is an urgent need for highly reliable, highly tuneable biomaterials and synthetic materials that are easy to produce in large amounts (scalable), ideally from detoxified agrofood wastes, to replace Matrigel using environmentally friendly processes and protocols. Cellular products with stable quality and functionality need to be produced in medium-large amounts to face the demand of cells for cell therapy transplantation linked to several disabling rare diseases, such as optic neuropathies, and diseases associated with age (with an expected increment of prevalence of 80% from 2030 to 2050). bioMAT4EYE proposal addresses the quest for new biomaterials based on polysaccharides of microbial and crustacean origin, all focused on promoting the differentiation of human induced pluripotent stem cells (hiPSCs) to functional and properly projecting retinal ganglion cells (RGCs), the neurons that selectively die in glaucoma and other optic neuropathies. Hence, we will optimize the microbial processes to target hydrogels with particular properties (rheology, interaction with hiPSCs, biodegradability), several surface coating procedures based on biopolymers, different conformational technologies to create 2D+ and 3D micro-bioreactors, operational conditions based on microfluidics and on-site/off-site sensors to assess the functionality (quality) of final RGCs.