Abstract
The blood-brain barrier (BBB) is a key biological interface that controls trafficking between the bloodstream and brain to maintain neural homeostasis. To carry out this role, the BBB exhibits several specialized properties, including limited permeability and active transporter function. These properties are often evaluated within in vitro BBB models, which can be utilized for high-throughput screening applications such as drug discovery. Here, we detail several common methods used to qualify in vitro BBB models, including measurement of transendothelial electrical resistance (TEER), determination of permeability coefficients (Pe) for small molecules, and assessment of efflux transporter activity. We describe these methods in the context of BBB endothelial cells derived from human-induced pluripotent stem cells (iPSCs), a model commonly employed in our research group.
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References
Abbott NJ, Rönnbäck L, Hansson E (2006) Astrocyte–endothelial interactions at the blood–brain barrier. Nat Rev Neurosci 7:41–53
Daneman R, Prat A (2015) The blood–brain barrier. Cold Spring Harb Perspect Biol 7:a020412
Obermeier B, Daneman R, Ransohoff RM (2013) Development, maintenance and disruption of the blood-brain barrier. Nat Med 19:1584–1596
Zlokovic BV (2008) The blood-brain barrier in health and chronic neurodegenerative disorders. Neuron 57:178–201
Montagne A, Nation DA, Pa J et al (2016) Brain imaging of neurovascular dysfunction in Alzheimer’s disease. Acta Neuropathol (Berl) 131:687–707
van de Haar HJ, Burgmans S, Jansen JFA et al (2016) Blood-brain barrier leakage in patients with early Alzheimer disease. Radiology 281:527–535
Yates PA, Desmond PM, Phal PM et al (2014) Incidence of cerebral microbleeds in preclinical Alzheimer disease. Neurology 82:1266–1273
Uetani H, Hirai T, Hashimoto M et al (2013) Prevalence and topography of small hypointense foci suggesting microbleeds on 3T susceptibility-weighted imaging in various types of dementia. Am J Neuroradiol 34:984–989
Simpson IA, Chundu KR, Davies-Hill T et al (1994) Decreased concentrations of GLUT1 and GLUT3 glucose transporters in the brains of patients with Alzheimer’s disease. Ann Neurol 35:546–551
Jagust WJ, Seab JP, Huesman RH et al (1991) Diminished glucose transport in Alzheimer’s disease: dynamic PET studies. J Cereb Blood Flow Metab 11:323–330
Hunt A, Schönknecht P, Henze M et al (2007) Reduced cerebral glucose metabolism in patients at risk for Alzheimer’s disease. Psychiatry Res Neuroimaging 155:147–154
Niwa K, Kazama K, Younkin SG et al (2002) Alterations in cerebral blood flow and glucose utilization in mice overexpressing the amyloid precursor protein. Neurobiol Dis 9:61–68
Deo AK, Borson S, Link JM et al (2014) Activity of P-glycoprotein, a β-amyloid transporter at the blood-brain barrier, is compromised in patients with mild Alzheimer disease. J Nucl Med 55:1106–1111
van Assema DM, Lubberink M et al (2012) Blood–brain barrier P-glycoprotein function in Alzheimer’s disease. Brain 135:181–189
Kortekaas R, Leenders KL, van Oostrom JCH et al (2005) Blood–brain barrier dysfunction in parkinsonian midbrain in vivo. Ann Neurol 57:176–179
Naik P, Cucullo L (2012) In vitro blood–brain barrier models: current and perspective technologies. J Pharm Sci 101:1337–1354
Dehouck M-P, Méresse S, Delorme P et al (1990) An easier, reproducible, and mass-production method to study the blood–brain barrier in vitro. J Neurochem 54:1798–1801
Franke H, Galla H-J, Beuckmann CT (1999) An improved low-permeability in vitro-model of the blood–brain barrier: transport studies on retinoids, sucrose, haloperidol, caffeine and mannitol. Brain Res 818:65–71
Patabendige A, Skinner RA, Abbott NJ (2013) Establishment of a simplified in vitro porcine blood–brain barrier model with high transendothelial electrical resistance. Brain Res 1521:1–15
Al Ahmad A, Taboada CB, Gassmann M et al (2011) Astrocytes and pericytes differentially modulate blood–brain barrier characteristics during development and hypoxic insult. J Cereb Blood Flow Metab 31:693–705
Al Ahmad A, Gassmann M, Ogunshola OO (2009) Maintaining blood–brain barrier integrity: pericytes perform better than astrocytes during prolonged oxygen deprivation. J Cell Physiol 218:612–622
Song L, Pachter JS (2003) Culture of murine brain microvascular endothelial cells that maintain expression and cytoskeletal association of tight junction-associated proteins. In Vitro Cell Dev Biol Anim 39:313–320
Smith QR, Rapoport SI (1986) Cerebrovascular permeability coefficients to sodium, potassium, and chloride. J Neurochem 46:1732–1742
Helms HC, Abbott NJ, Burek M et al (2016) In vitro models of the blood–brain barrier: an overview of commonly used brain endothelial cell culture models and guidelines for their use. J Cereb Blood Flow Metab 36:862–890
Pardridge WM (1998) CNS drug design based on principles of blood-brain barrier transport. J Neurochem 70:1781–1792
Syvänen S, Lindhe Ö, Palner M et al (2009) Species differences in blood-brain barrier transport of three positron emission tomography radioligands with emphasis on P-glycoprotein transport. Drug Metab Dispos 37:635–643
Weksler BB, Subileau EA, Perrière N et al (2005) Blood-brain barrier-specific properties of a human adult brain endothelial cell line. FASEB J 19:1872–1874
Lippmann ES, Azarin SM, Kay JE et al (2012) Derivation of blood-brain barrier endothelial cells from human pluripotent stem cells. Nat Biotechnol 30:783–791
Lippmann ES, Al-Ahmad A, Azarin SM et al (2014) A retinoic acid-enhanced, multicellular human blood-brain barrier model derived from stem cell sources. Sci Rep 4:4160
Wilson HK, Canfield SG, Hjortness MK et al (2015) Exploring the effects of cell seeding density on the differentiation of human pluripotent stem cells to brain microvascular endothelial cells. Fluids Barriers CNS 12:13
Hollmann EK, Bailey AK, Potharazu AV et al (2017) Accelerated differentiation of human induced pluripotent stem cells to blood–brain barrier endothelial cells. Fluids Barriers CNS 14:9
Acknowledgments
Our research efforts in this area are supported by a NARSAD Young Investigator Award from the Brain and Behavior Research Foundation (ESL) and grant A20170945 from the Alzheimer’s Disease Research Program through the BrightFocus Foundation (ESL). EHN is supported by a National Science Foundation Graduate Research Fellowship.
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Neal, E.H., Shi, Y., Lippmann, E.S. (2019). In Vitro Blood-Brain Barrier Functional Assays in a Human iPSC-Based Model. In: Aschner, M., Costa, L. (eds) Cell Culture Techniques. Neuromethods, vol 145. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9228-7_1
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DOI: https://doi.org/10.1007/978-1-4939-9228-7_1
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