Protocol to analyze the transmigration efficiency of T. brucei using an in vitro model of the blood-cerebrospinal fluid barrier

STAR Protoc. 2022 Dec 16;3(4):101676. doi: 10.1016/j.xpro.2022.101676. Epub 2022 Sep 13.

Abstract

At present, the only approach to investigate the transmigration of Trypanosoma brucei, the causative agent of human African trypanosomiasis, from blood to cerebrospinal fluid is through animal experiments. This protocol details how to analyze the transmigration efficiency using an in vitro model of the blood-cerebrospinal fluid (blood-CSF) barrier. We describe how to grow human choroid plexus epithelial cells on cell culture filter inserts to form the barrier, followed by isolating and quantifying genomic DNA of transmigrated parasites by qPCR. For complete details on the use and execution of this protocol, please refer to Speidel et al. (2022).

Keywords: Cell biology; Cell culture; Cell-based assays; Health sciences; Microbiology; Model organisms; Molecular biology; Neuroscience.

MeSH terms

  • Animals
  • Blood-Brain Barrier*
  • Cell Culture Techniques
  • Epithelial Cells*
  • Humans