Isolation and culture of adult mouse vestibular nucleus neurons

Abstract

Background/aim: Isolated cell cultures are widely used to study neuronal properties due to their advantages. Although embryonic animals are preferred for culturing, their morphological or electrophysiological properties may not reflect adult neurons, which may be important in neurodegenerative diseases. This paper aims to develop a method for preparing isolated cell cultures of medial vestibular nucleus (MVN) from adult mice and describe its morphological and electrophysiological properties.Materials and methods: Vestibular nucleus neurons were mechanically and enzymatically isolated and cultured using a defined medium with known growth factors. Cell survival was measured with propidium iodide, and electrophysiological properties were investigated with current-clamp recording.Results: Vestibular neurons grew neurites in cultures, gaining adult-like morphological properties, and stayed viable for 3 days in culture. Adding bovine calf serum, nerve growth factor, or insulin-like growth factor into the culture medium enhanced neuronal viability. Current-clamp recording of the cultured neurons revealed tonic and phasic-type neurons with similar input resistance, resting membrane potential, action potential amplitude, and duration. Conclusion:Vestibular neurons from adult mice can be cultured, and regenerate axons in a medium containing appropriate growth factors. Culturing adult vestibular neurons provides a new method to study age-related pathologies of the vestibular system.

Modulation of the intrinsic neuronal excitability by multifunctional liposomes tailored for the treatment of Alzheimer's disease

“…In the literature, there is only one paper aiming at developing a method for preparing isolated cells of medial vestibular nucleus (MVN) from adult mice, which however remain viable only for 3 days in culture. 29 These cells, however, are not consistent with our model. Moreover, in order to study the electrical properties of isolated neurons, embryonic or neonatal animals are preferred for culturing.…”

Analysis of isolation of cerebral cortical neurons in rats by different methods

“…The separation and culture of neuronal cells from the central nervous system are essential for the study of neuropharmacology, neurodegenerative diseases, and nerve regeneration mechanisms. Common methods for obtaining single-cell suspensions of neurons are the grinding method and the collagenase and trypsin methods (Him et al, 2017;Katzenell et al, 2017;Leong et al, 2013;Yu et al, 2016;Zhang and Hu, 2013), and the grinding method are applied to extract and separate astrocytes to study the effect of ADPribosylation on the aging rat brain (Manoochehr and Azadeh, 2019) and the neural stem cells were extracted and separated to study the Gene expression profile of Sox1, Sox2, p53, Bax and Nestin in adult mouse brain tissues (Wang et al, 2019), but it is still not clear which method can quickly and efficiently obtain neuronal cells from the adult rat brain. Although embryonic neurons are a simple and rapid source of primary neuronal cells, their developmental stages are not suitable for studying many neurodegenerative diseases that occur in later life.…”