Introduction
In vitro systems have had an important role in the development of our current understanding of the biochemical properties of the blood–brain barrier (BBB) (Joo, 1985, 1992, 1993). In particular, the development of brain microvessel endothelial cell (BMEC) culture systems, which retain many of the BBB properties in vitro, has provided simple, dynamic experimental systems to evaluate and elaborate on the biochemical and molecular basis of the BBB (Laterra and Goldstein, 1993). This chapter focuses on some of the current methodologies for the establishment of primary cultures of BMECs, BMEC and astrocyte co-culture systems, and the configurations available for studying BMEC transport processes.
Cell culture methods
Essentially three types of BMEC cultures are currently used by researchers. These include the primary cultures, co-culture systems, and cell lines. Each has demonstrated and has contributed basic information on cellular, biochemical, and molecular properties of the BBB (Joo, 1985, 1992, 1993; Laterra and Goldstein, 1993). For each system, we have provided a summary of methodologies involved for establishment and some comments on the advantages and disadvantages of the culture type. For a given researcher, choice of these systems will generally be dictated by the specific BBB application or parameter one wants to investigate.