The choroid plexus epithelium forms the interface between the blood and the cerebrospinal fluid. In addition to its barrier
function resulting from the presence of tight junctions sealing the epithelial cells together, the choroid plexus epithelium
fulfills vectorial transport (influx and efflux), neuroprotective, antioxidant and secretory functions, all relevant to different
aspects of neurotoxicological sciences. To investigate these choroidal functions without the interference of the blood–brain
barrier proper and brain parenchyma, in vitro cellular models of the blood–cerebrospinal fluid barrier, retaining the differentiated
phenotype of the choroidal epithelium, have been established, taking advantage of the advent of refined culture methods and
availability of permeable membranes. This chapter provides information to help investigators to set up and characterize choroid
plexus epithelial cells in culture in bicameral devices. It first describes the factors that are critical to isolate the cells
and select the culture conditions, and provides a survey of available cell lines with their advantages and limitations. Then
the primordial specific choroidal features that can be examined within a validation scheme are discussed, emphasizing the
need for a careful interpretation of their significance. In a third part, selected examples of studies performed with these
models are presented, highlighting their potential applications in the field of neurotoxicology.