The area surrounding the central canal of the mammalian spinal cord is known to contain two cell types: ependymal cells and cerebrospinal fluid contacting cells (CSFcCs). It has been defined as a neurogenic niche due to the presence of a subpopulation of ependymal cells that have characteristics of neural stem cells and the presence of CSFcCs that appear to be immature neurones. To date the function and characteristics of both these cell types are poorly understood, this needs to be rectified if we are to comprehend how the neurogenic niche functions. This study used electrophysiology and immunohistochemistry to characterise ependymal cells and CSFcCs. For whole cell patch clamp electrophysiology, Wistar rats or C57/b6 mice (P11-21) were anaesthetised by I.P administration of sodium pentobarbitone (60 mg/kg), the spinal cords dissected and 300 µm slices cut. Recordings were made in current clamp, whilst dyes were added intracellularly. Recordings revealed that ependymal cells were a uniform population with a resting membrane potential of -75.7 mV ± 0.7 (SE, n = 63), an input resistance of 100.1 MΩ ± 15.2 (n = 63) and no spontaneous or evoked activity. Dyes revealed a subpopulation of cells that possessed dorsally projecting fibres. Based on electrophysiology, the CSFcCs were categorised into four subtypes: subtype 1 (n = 7) had similar electrophysiological properties to ependymal cells but were morphologically defined as CSFcCs, subtypes 2 (n = 19), 3 (n = 95) and 4 (n = 44) were defined by their ability to produce a depolarisation, a spike or 2 or more spikes, respectively, in response to the injection of positive current. For immunohistochemistry, adult C57/b6 mice were anaesthetised by I.P administration of sodium pentobarbitone (60mg/kg), perfused with 4 % paraformaldehyde and the spinal cord was removed and sectioned. CSFcCs were identified by rabbit anti-PKD2L1. Rat anti-5HT revealed that 58 ± 17 % of CSFcCs had 5HT-positive terminals in close apposition and rat anti-substance P revealed that 52 ± 7 % of CSFcCs had substance P-positive terminals in close apposition. Electrophysiology, however, showed that neither 5HT (n = 4, mouse; n = 9, rat) nor substance P (n =12, rat) had an effect on the input resistance or membrane potential of CSFcCs. As positive controls, CSFcCs were shown to respond to GABA (n = 4, mouse; n = 17, rat) and nearby interneurones in lamina X were shown to respond to 5HT (n = 3, rat) and substance P (n = 4, rat). The results to date are only the beginning of the characterisation of these cells; this must be built on to determine whether CSFcCs are capable of maturing and integrating with existing circuits. Understanding this area to the point where we can manipulate both ependymal cells and CSFcCs could lead to beneficial treatments for numerous spinal cord pathologies.