Group B Streptococcus (GBS) is a human pathogen and a leading cause of neonatal meningitis, pneumonia, and sepsis in newborns and adults. Group B Streptococcal meningitis is caused when blood-borne GBS interacts with and disrupts the blood-brain barrier, causing inflammation. The blood-brain barrier (BBB) is comprised of highly specialized brain endothelial cells (BECs) that serve as the interface between the circulation and the central nervous system (CNS), creating a tight endothelial cell barrier. Modeling the BBB using an induced pluripotent stem-cell (iPSC)-derived brain-like endothelial cell model (iBEC) allows for robust expression and retention of BBB phenotypes when compared to other in vitro models. This chapter describes the derivation of iBECs, the culture of GBS, and the use of various molecular biology techniques to closely investigate the host–pathogen interactions between the blood-brain barrier and Group B Streptococcus.