In Vitro Visualization of Cell-to-Cell Interactions Between Natural Killer Cells and Sensory Neurons

Abstract

Cell-to-cell interactions between the immune and nervous systems are increasingly recognized for their importance in health and disease. Assessment of cellular neuro-immune interactions can be aided by coculture of two (or more) cells in an in vitro model system that preserves the morphology of neuronal cells. Here we describe methods to investigate the cytotoxic effector functions of natural killer cells on sensory neurons isolated from syngeneic embryonic and adult mice. We present methods for the morphological analysis of axon fragmentation (pruning) and dynamic cell function via live confocal calcium imaging. These techniques can easily be adapted to study interactions between other combinations of immune cell subsets and neuronal populations.

Endogenous Glycoprotein GPM6a Is Involved in Neurite Outgrowth in Rat Dorsal Root Ganglion Neurons

“…HBSS was carefully aspirated and replaced with a pre-warmed collagenase type 1A solution (1 mL per 6 isolated DRGs) and incubated 30 min at 37 °C. Then, an additional digestion for 30 min in 1 mL of 0.25% trypsin in HBSS was carried out at 37 °C according to Kim et al [ 33 ]. The supernatant was aspirated, and the DRG pellet was resuspended in adult growth medium (AGM, Ham’s F12 medium (GIBCO, Thermo Fisher Scientific, Waltham, MA, USA) containing 10% horse serum, 2 mM L-glutamine, and 50 U/mL penicillin/streptomycin.…”