Drug delivery across the blood-brain barrier (BBB) is a major obstacle for the development of effective treatments of many central nervous system disorders. Sophisticated cell culture models of the BBB have helped us to identify, characterize, and validate a novel targeted drug delivery technology, designated 2B-Trans, for the receptor-mediated uptake and transport of drugs across the BBB. This paper describes in great detail how such a BBB cell culture model should be prepared and handled, and applied for the use of targeted drug delivery across the BBB.