An in vitro Blood-brain Barrier Model to Study the Penetration of Nanoparticles
Sezin Aday,
Wen Li,
Jeffrey Karp,
Nitin Joshi
Affiliations
Sezin Aday
Center for Nanomedicine, Department of Anesthesiology, Massachusetts Institute of Technology, Boston, MA 02115, USADepartment of Chemical Engineering and Koch Institute for Integrative Cancer Research, Brigham and Women's Hospital, Cambridge, MA 02139, USA, Harvard Medical School, Boston, MA 02115, USA
Wen Li
Center for Nanomedicine, Department of Anesthesiology, Perioperative and Pain Medicine, Brigham and Women’s Hospital, Boston, MA 02115, USADepartment of Chemical Engineering and Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA, Harvard Medical School, Boston, MA 02115, USA
Jeffrey Karp
Center for Nanomedicine, Department of Anesthesiology, Perioperative and Pain Medicine, Brigham and Women’s Hospital, Boston, MA 02115, USAHarvard Medical School, Boston, MA 02115, USA, Harvard–Massachusetts Institute of Technology Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA, Broad Institute, Cambridge, MA 02142, USA, Harvard Stem Cell Institute, Cambridge, MA 02138, USA
Nitin Joshi
Center for Nanomedicine, Department of Anesthesiology, Perioperative and Pain Medicine, Brigham and Women’s Hospital, Boston, MA 02115, USADepartment of Chemical Engineering and Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA, Harvard Medical School, Boston, MA 02115, USA
The blood-brain barrier (BBB), a crucial protection mechanism in the central nervous system (CNS), is a selective barrier comprised of endothelial cells. It hampers the development of therapeutic and diagnostic tools for neurological diseases due to the poor penetration of most of these agents. Rationally engineered nanoparticles (NP) can facilitate the transport of therapeutic and diagnostic agents across the BBB. However, evaluating BBB penetration by NP majorly relies on the use of expensive and time-consuming animal experiments with low throughput. In vitro BBB models composed of brain endothelial cells can be a useful tool to rapidly screen multiple NP formulations to compare their BBB penetration ability and identify optimal formulations for in vivo validation. In this protocol, we present an in vitro model of BBB developed using murine cerebral cortex endothelial cells (bEnd.3). bEnd.3 is a commercially available, easy to manipulate cell line that forms tight junctions with potent paracellular barrier property. The protocol includes culturing of bEnd.3 cells, establishment of the in vitro model, and assessing NP permeability. We believe that, due to its simplicity and consistency, this step-by-step protocol can be easily used by researchers to screen NP-based drug delivery systems for BBB penetration.Graphic abstract:
