A preliminary in vitro study on Adult Neural Stem cells from 3xTg mice

Abstract

AbstractBackgroundIn AD, abnormalities in brain wiring and variations of the brain energetic metabolism are considered a key pathological framework of cognitive decline (Heininger, 2000). Neural stem cells (NSCs) can contribute to brain regeneration and, their transplantation improves the behaviour of neurodegenerative mice models (De Gioia et.al., 2021) although, the molecular mechanisms involved are still unidentified. Our preliminary study aims to explore the potential differences between NSCs‐derived neurons from 6‐months‐old healthy and AD mice in terms of neuronal network morphology. Furthermore, the study wants to provide a tool to investigate whether cell energy metabolism is modified, considering that bioenergetic failure can influence the regenerative potential of adult AD brain, an issue that is still ill defined.MethodNSCs were isolated from subventricular zone of healthy (H) and 3xTg (AD) mice. NSCs selection was based on their ability to proliferate and to form 3D floating neurospheres. Isolated cells were amplified and characterised for their expression of stem‐related markers. Subsequently, neuron and astrocyte differentiation protocols were set up. Morphological analyses were performed on H and AD NSC‐derived neurons and the expressions of glutamate transporters and sodium/calcium exchangers (NCXs) were investigated.ResultH and AD NSCs were strongly positive for Nestin and Sox2 stem markers and a faint expression of NeuN and Gfap, markers related to neuron and astrocyte mature phenotype respectively, was detected. These data confirmed the purity of the cell cultures as well as the reliability of the protocols. Morphological investigations highlighted a different distribution of the neurite network between NSCs‐derived neurons isolated from H and AD mice. Furthermore, H and AD mature cells expressed glutamate transporters essential not only for cellular transmission but also for energy production by their interplay with NCXs.ConclusionPreliminary data suggest that the pathological features of adult neurons from AD brain can be traced back to “stem level” and continue during differentiation. The different morphological organization of neurite network observed in NSCs‐derive AD neurons could be involved in the alteration of the impulse transmission and plasticity. Further studies will be necessary to better clarify the consequence of dysregulated glutamate transporters as well the role of astrocytes.