3D in vitro models for neural progenitor cell differentiation and network formation
[eng] Conventional 2D in vitro and animal models have been found inadequate to fully uncover the intricate mechanisms happening in the brain and its diseases. Engineering-based models emerge as promising alternatives to the development of more complex and dependable brain models. Scaffold-based cult...
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| Tipo de recurso: | tesis doctoral |
| Estado: | Versión publicada |
| Fecha de publicación: | 2023 |
| País: | España |
| Institución: | Universidad de Barcelona |
| Repositorio: | Dipòsit Digital de la UB |
| OAI Identifier: | oai:diposit.ub.edu:2445/203343 |
| Acceso en línea: | https://hdl.handle.net/2445/203343 http://hdl.handle.net/10803/689241 |
| Access Level: | acceso abierto |
| Palabra clave: | Impressió 3D Materials biomèdics Extrusió (Mecànica) Microfluídica Diferenciació cel·lular Cèl·lules mare Three-dimensional printing Biomedical materials Extrusion process Microfluidics Cell diferentiation Stem cells |
| Sumario: | [eng] Conventional 2D in vitro and animal models have been found inadequate to fully uncover the intricate mechanisms happening in the brain and its diseases. Engineering-based models emerge as promising alternatives to the development of more complex and dependable brain models. Scaffold-based culture is one example of this type of models where cells are cultured in a biomaterial that simulates more closely the organ environment and function. Another example is microfluidic devices as they combine micropatterned platforms with cell culture to create models with a tuneable framework using small amounts of resources. In this work, we used methacrylated gelatine, methacrylated alginate and hyaluronic acid to develop a biomaterial with adjustable mechanical properties and biocompatibility that resembles the extracellular matrix for neural culture. This composite biomaterial presented suitable physical properties with high water intake, low stiffness, and slow degradation. Through the evaluation of cell viability and the expression of differentiation markers, we could observe that our formulation was compatible with the culture, differentiation, and network formation of murine neuroprogenitor cells into early neurons. Calcium imaging also validated the activity of the cells in the system. This biomaterial was assessed as bioink for the extrusion bioprinting of the same cell line, presenting good definition, high cell viability, the expression of differentiation markers, and functional activity. The combination of our biomaterial with a 3D microfluidic device of three parallel channels separated by triangular-shaped pillars was also possible, with viable cultures up to 21 days. We complemented these results with the evaluation of the compatibility of our hydrogel with human induced pluripotent stem cells and observed good viability results and the start of differentiation into dopaminergic neurons. In parallel to the development of our biomaterial, we designed a microfluidic device capable of being used for the culture of different neuronal subtypes. The design consisted of three parallel channels connected through obliquus microchannels with dimensions below 5 µm to isolate and direct axons from the lateral channels to the central channel. This model was used to recreate the cortico-striatal circuit with successful differentiation of cortical and striatal cells and the influence of the co-culture was validated through calcium imaging. The dopaminergic-striatal circuit was also modelled using our device with results pointing to the influence of the striatal neurons in the differentiation of dopaminergic neurons. |
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