Effects of Different Laminins on Human Neural Progenitor Cells Cultured in 3D Electrospun Fibers
Abstract: A low success rate of drug candidates in clinical trials, and especially in brain research, motivates development of better screening models. Hence, in neuroscience and many other fields of Life Science, development of more physiological relevant experimental cell-based models is motivated. It’s believed that more in vivo-like models will give a better outcome in drug discovery- and development. However, until today, cell models typically include non-human cells, a two-dimensional (2D) culture substrate, which may not include proteins present in the cells’ native extracellular matrix (ECM). The aim of this thesis work is, therefore, to explore the potential to generate a more physiological relevant cell-based model for brain research, by using a human brain cell line, three-dimensional (3D) culture substrates and the ECM protein laminin. Electrospinning was used to fabricate 3D culture fiber scaffolds. A human neural progenitor cell (hNPC) line, with demonstrated capacity to form neurons was used and their innate laminin production at 0 and 20 days in vitro (DIV) were characterized. 2D and 3D substrates were coated with one subtype of mouse laminin, six subtypes of human recombinant laminins and non-coated substrates served as control. Bio- and immunochemical assays was used to analyze cell viability and overall and neuronal differentiation potential. We revealed expression of a variety of laminin chains in hNPC, both in an immature stage and after 20 DIV. Compared to the non-coated groups, cell viability was in general equal or better in all laminin-treated groups, especially for the human biolaminins. Level of differentiation stage, judged by nestin (marker for NPC)- and βIII-tubulin (marker for early neurons) expression in immuno-labeling and western blot, may be affected by different human biolaminins. Specifically, the human biolaminins seem to promote neuronal differentiation compared to the mouse counterpart. Our initial results demonstrate that brain cells have the potential to survive well in functionalized 3D scaffolds, and that different human laminin subtypes most likely affect their viability, differentiation potential differently. However, more studies are needed to confirm the effect of specific human laminins and before a protocol for the culture of in vivo-like human in vitro brain models can be presented.
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