Methods for shipping live primary cortical and hippocampal neuron cultures from postnatal mice

Abstract

Primary neuronal cultures have proven to be a powerful tool for studying mechanisms in neuroscience. It is technically challenging and expensive to reproduce high quality viable neuronal cultures. Laboratories that are not experienced or equipped to prepare primary neuron cultures may have difficulty producing consistent cultures for experiments. It has previously been shown that live rat embryonic hippocampal cultures can be shipped from laboratories that produce them. Here, we show that variations to this procedure allow for shipping postnatal mouse cultures of hippocampal and cortical primary neurons using standard commercial couriers. We also show that after shipping, primary neurons are viable, express synaptic markers, and demonstrate physiological activity, making them relevant models over immortalized cell lines. Among the many applications of this technique would be the preparation of cultured neurons from transgenic mouse lines in one laboratory and sharing them with distant collaborators, reducing variability.

Keywords: Electrophysiology; Methodology; Primary neuronal culture; Shipping.

Graphical abstract

Fig. 1

Identity and viability of mouse neurons after shipment. A. Expression of neuron-specific IIIβ-tubulin (IIIβ-Tub) and the synaptic markers synapsin 1 (Syn1) in 14 DIV neurons following a shipment. B. Expression of IIIβ-Tub and a marker of GABAergic synaptic terminals (VGAT, vesicular GABA transporter) in 14 DIV neurons. C. Spontaneous post synaptic currents (sPSCs) recorded from 14 DIV mouse neurons. D. Induced action potential firing of 14 DIV neurons. E. PI staining of mouse neurons prior to shipment (2 DIV), after shipment (4 DIV), and non-shipped conditions (4 DIV). All data are presented as means ± SD. Significant differences to control are represented as *, p < 0.05, as determined by Two-way ANOVA. N = 6.