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Cited by 42Barrier Structure and Function StudiesDrug Transport and Resistance MechanismsExtracellular vesicles in disease

Current Status of In vitro Models of the Blood-brain Barrier

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Dong, Xiaowei, Shah, Brijesh·2022·Current Drug Delivery
DOI 10.2174/1567201819666220303102614PMC PMC9440157
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Abstract

Disorders of the brain constitute the most debilitating situation globally with increased mortality rates every year, while brain physiology and cumbersome drug development processes exacerbate this. Although blood-brain barrier (BBB) and its components are important for brain protection, their complexity creates major obstacles for brain drug delivery, and the BBB is the primary cause of treatment failure, leading to disease progression. Therefore, developing an ideal platform that can predict the behavior of a drug delivery system in the brain at the early development phase is extremely crucial. In this direction, in the last two decades, numerous in vitro BBB models have been developed and investigated by researchers to understand the barrier properties and how closely the in vitro models mimic in vivo BBB. In-vitro BBB models mainly involve the culture of endothelial cells or their coculture with other perivascular cells either in two or three-dimensional platforms. In this article, we have briefly summarized the fundamentals of BBB and outlined different types of in vitro BBB models with their pros and cons. Based on the available reports, no model seems to be robust that can truly mimic the entire properties of the in vivo BBB microvasculature. However, human stem cells, coculture and threedimensional models have been found to mimic the complexity of the barrier integrity not completely but more precisely than other in vitro models. More studies aiming towards combining these models together would be needed to develop an ideal in vitro model that can overcome the existing limitations and unravel the mysterious BBB vasculature.

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  • BBB transwell models allow transport measurements but are low-throughput and do not capture transfection.

    “BBB transwell models have a semipermeable membrane separating two compartments allowing for transport measurements but are low-throughput as they are typically only used in 12- or 24-well formats and do not capture transfection.30,31”

    From Predictive High-Throughput Platform for Dual Screening of mRNA Lipid Nanoparticle Blood–Brain Barrier Transfection and Crossing (2024) · cites this paper as [30]

    The Monolayer Models section says transwell chambers have a semipermeable membrane separating two sides and allowing solute exchange, supporting transport measurements. It describes transwell models as scalable rather than low-throughput, and our copy does not support the claimed 12- or 24-well restriction or say that they cannot capture transfection (page numbers unavailable). (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • Brain endothelial monolayers grown in traditional 96-well plates can screen for transfection in a high-throughput manner but cannot measure transport across the blood-brain barrier.

    “Brain endothelial monolayers grown in traditional 96-well plates can screen for transfection in a high-throughput manner, but the lack of two compartments means that they cannot measure transport across the BBB.30”

    From Predictive High-Throughput Platform for Dual Screening of mRNA Lipid Nanoparticle Blood–Brain Barrier Transfection and Crossing (2024) · cites this paper as [30]

    The Monolayer Models section says monocultures in Petri dishes lack separate blood and brain compartments and are unsuitable for studying drug transport across the BBB. It does not identify traditional 96-well plates or say these cultures can screen for transfection in a high-throughput manner. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

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Research works printed but not linked: 59

  1. [1]Wilhelm I; Fazakas C; Krizbai IA, In vitro models of the blood-brain barrier. Acta Neurobiol Exp (Wars) 2011, 71 (1), 113– 28. 21499332
  2. [2]Pardridge WM, Blood-brain barrier delivery. Drug Discov Today 2007, 12 (1–2), 54– 61. 17198973
  3. [3]Jamieson JJ; Searson PC; Gerecht S, Engineering the human blood-brain barrier in vitro. J Biol Eng 2017, 11, 37. 29213304 PMC5713119
  4. [4]Wilhelm I; Krizbai IA, In vitro models of the blood-brain barrier for the study of drug delivery to the brain. Mol Pharm 2014, 11 (7), 1949– 63. 24641309
  5. [6]Bagchi S; Chhibber T; Lahooti B; Verma A; Borse V; Jayant RD, In-vitro blood-brain barrier models for drug screening and permeation studies: an overview. Drug Des Devel Ther 2019, 13, 3591– 3605. PMC6805046 31695329
  6. [8]Crone C; Christensen O, Electrical resistance of a capillary endothelium. J Gen Physiol 1981, 77 (4), 349– 71. 7241087 PMC2215420
  7. [9]Olesen SP; Crone C, Electrical resistance of muscle capillary endothelium. Biophys J 1983, 42 (1), 31– 41. 6601500 PMC1329200
  8. [10]Wolff A; Antfolk M; Brodin B; Tenje M, In Vitro Blood-Brain Barrier Models-An Overview of Established Models and New Microfluidic Approaches. J Pharm Sci 2015, 104 (9), 2727– 46. 25630899
  9. [12]Weksler BB; Subileau EA; Perriere N; Charneau P; Holloway K; Leveque M; Tricoire-Leignel H; Nicotra A; Bourdoulous S; Turowski P; Male DK; Roux F; Greenwood J; Romero IA; Couraud PO, Blood-brain barrier-specific properties of a human adult brain endothelial cell line. FASEB J 2005, 19 (13), 1872– 4. 16141364
  10. [13]Myers JS; Hare J; Sang QA, A Simple Adaptable Blood-Brain Barrier Cell Model for Screening Matrix Metalloproteinase Inhibitor Functionality. Methods Mol Biol 2017, 1579, 287– 296. 28299744
  11. [14]Czupalla CJ; Liebner S; Devraj K, In vitro models of the blood-brain barrier. Methods Mol Biol 2014, 1135, 415– 37. 24510883
  12. [15]Dehouck MP; Jolliet-Riant P; Bree F; Fruchart JC; Cecchelli R; Tillement JP, Drug transfer across the blood-brain barrier: correlation between in vitro and in vivo models. J Neurochem 1992, 58 (5), 1790– 7. 1560234
  13. [16]Andjelkovic AV; Stamatovic SM; Phillips CM; Martinez-Revollar G; Keep RF, Modeling blood-brain barrier pathology in cerebrovascular disease in vitro: current and future paradigms. Fluids Barriers CNS 2020, 17 (1), 44. 32677965 PMC7367394
  14. [17]Sivandzade F; Cucullo L, In-vitro blood-brain barrier modeling: A review of modern and fast-advancing technologies. J Cereb Blood Flow Metab 2018, 38 (10), 1667– 1681. 30058456 PMC6168917
  15. [18]Bagchi S; Lahooti B; Chhibber T; Varahachalam SP; Mittal R; Joshi A; Jayant RD, In Vitro Models of Central Nervous System Barriers for Blood-Brain Barrier Permeation Studies. In Nanomedicines for Brain Drug Delivery, Morales JO; Gaillard PJ, Eds. Humana: New York, 2020; Vol. 157, pp 235– 253.
  16. [19]Hartmann C; Zozulya A; Wegener J; Galla HJ, The impact of glia-derived extracellular matrices on the barrier function of cerebral endothelial cells: an in vitro study. Exp Cell Res 2007, 313 (7), 1318– 25. 17346702
  17. [20]Helms HC; Abbott NJ; Burek M; Cecchelli R; Couraud PO; Deli MA; Forster C; Galla HJ; Romero IA; Shusta EV; Stebbins MJ; Vandenhaute E; Weksler B; Brodin B, 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 2016, 36 (5), 862– 90. 26868179 PMC4853841
  18. [21]DeBault LE; Cancilla PA, gamma-Glutamyl transpeptidase in isolated brain endothelial cells: induction by glial cells in vitro. Science 1980, 207 (4431), 653– 5. 6101511
  19. [22]Gaillard PJ; Voorwinden LH; Nielsen JL; Ivanov A; Atsumi R; Engman H; Ringbom C; de Boer AG; Breimer DD, Establishment and functional characterization of an in vitro model of the blood-brain barrier, comprising a co-culture of brain capillary endothelial cells and astrocytes. Eur J Pharm Sci 2001, 12 (3), 215– 22. 11113640
  20. [23]Jeliazkova-Mecheva VV; Bobilya DJ, A porcine astrocyte/endothelial cell co-culture model of the blood-brain barrier. Brain Res Brain Res Protoc 2003, 12 (2), 91– 8. 14613810
  21. [24]Nakagawa S; Deli MA; Nakao S; Honda M; Hayashi K; Nakaoke R; Kataoka Y; Niwa M, Pericytes from brain microvessels strengthen the barrier integrity in primary cultures of rat brain endothelial cells. Cell Mol Neurobiol 2007, 27 (6), 687– 94. 17823866 PMC11517186
  22. [25]Dohgu S; Takata F; Yamauchi A; Nakagawa S; Egawa T; Naito M; Tsuruo T; Sawada Y; Niwa M; Kataoka Y, Brain pericytes contribute to the induction and up-regulation of blood-brain barrier functions through transforming growth factor-beta production. Brain Res 2005, 1038 (2), 208– 15. 15757636
  23. [26]Nakagawa S; Deli MA; Kawaguchi H; Shimizudani T; Shimono T; Kittel A; Tanaka K; Niwa M, A new blood-brain barrier model using primary rat brain endothelial cells, pericytes and astrocytes. Neurochem Int 2009, 54 (3–4), 253– 63. 19111869
  24. [27]Canfield SG; Stebbins MJ; Morales BS; Asai SW; Vatine GD; Svendsen CN; Palecek SP; Shusta EV, An isogenic blood-brain barrier model comprising brain endothelial cells, astrocytes, and neurons derived from human induced pluripotent stem cells. J Neurochem 2017, 140 (6), 874– 888. 27935037 PMC5339046
  25. [28]Yamamizu K; Iwasaki M; Takakubo H; Sakamoto T; Ikuno T; Miyoshi M; Kondo T; Nakao Y; Nakagawa M; Inoue H; Yamashita JK, In Vitro Modeling of Blood-Brain Barrier with Human iPSC-Derived Endothelial Cells, Pericytes, Neurons, and Astrocytes via Notch Signaling. Stem Cell Reports 2017, 8 (3), 634– 647. 28238797 PMC5355679
  26. [29]Mantle JL; Min L; Lee KH, Minimum Transendothelial Electrical Resistance Thresholds for the Study of Small and Large Molecule Drug Transport in a Human in Vitro Blood-Brain Barrier Model. Mol Pharm 2016, 13 (12), 4191– 4198. 27934481
  27. [30]Kokubu Y; Yamaguchi T; Kawabata K, In vitro model of cerebral ischemia by using brain microvascular endothelial cells derived from human induced pluripotent stem cells. Biochem Biophys Res Commun 2017, 486 (2), 577– 583. 28336435
  28. [31]Adriani G; Ma D; Pavesi A; Kamm RD; Goh EL, A 3D neurovascular microfluidic model consisting of neurons, astrocytes and cerebral endothelial cells as a blood-brain barrier. Lab Chip 2017, 17 (3), 448– 459. 28001148
  29. [32]Crisan M; Yap S; Casteilla L; Chen CW; Corselli M; Park TS; Andriolo G; Sun B; Zheng B; Zhang L; Norotte C; Teng PN; Traas J; Schugar R; Deasy BM; Badylak S; Buhring HJ; Giacobino JP; Lazzari L; Huard J; Peault B, A perivascular origin for mesenchymal stem cells in multiple human organs. Cell Stem Cell 2008, 3 (3), 301– 13. 18786417
  30. [33]Tian X; Brookes O; Battaglia G, Pericytes from Mesenchymal Stem Cells as a model for the blood-brain barrier. Sci Rep 2017, 7, 39676. 28098158 PMC5241806
  31. [34]Cucullo L; Couraud PO; Weksler B; Romero IA; Hossain M; Rapp E; Janigro D, Immortalized human brain endothelial cells and flow-based vascular modeling: a marriage of convenience for rational neurovascular studies. J Cereb Blood Flow Metab 2008, 28 (2), 312– 28. 17609686
  32. [35]Cucullo L; Marchi N; Hossain M; Janigro D, A dynamic in vitro BBB model for the study of immune cell trafficking into the central nervous system. J Cereb Blood Flow Metab 2011, 31 (2), 767– 77. 20842162 PMC3049530
  33. [36]Siddharthan V; Kim YV; Liu S; Kim KS, Human astrocytes/astrocyte-conditioned medium and shear stress enhance the barrier properties of human brain microvascular endothelial cells. Brain Res 2007, 1147, 39– 50. 17368578 PMC2691862
  34. [37]Prabhakarpandian B; Shen MC; Nichols JB; Mills IR; Sidoryk-Wegrzynowicz M; Aschner M; Pant K, SyM-BBB: a microfluidic Blood Brain Barrier model. Lab Chip 2013, 13 (6), 1093– 101. 23344641 PMC3613157
  35. [38]Deosarkar SP; Prabhakarpandian B; Wang B; Sheffield JB; Krynska B; Kiani MF, A Novel Dynamic Neonatal Blood-Brain Barrier on a Chip. PLoS One 2015, 10 (11), e0142725. 26555149 PMC4640840
  36. [39]Yeon JH; Na D; Choi K; Ryu SW; Choi C; Park JK, Reliable permeability assay system in a microfluidic device mimicking cerebral vasculatures. Biomed Microdevices 2012, 14 (6), 1141– 8. 22821236
  37. [40]Oddo A; Peng B; Tong Z; Wei Y; Tong WY; Thissen H; Voelcker NH, Advances in Microfluidic Blood-Brain Barrier (BBB) Models. Trends Biotechnol 2019, 37 (12), 1295– 1314. 31130308
  38. [41]Jiang L; Li S; Zheng J; Li Y; Huang H, Recent Progress in Microfluidic Models of the Blood-Brain Barrier. Micromachines (Basel) 2019, 10 (6). PMC6630552 31195652
  39. [43]Kim JA; Kim HN; Im SK; Chung S; Kang JY; Choi N, Collagen-based brain microvasculature model in vitro using three-dimensional printed template. Biomicrofluidics 2015, 9 (2), 024115. 25945141 PMC4401807
  40. [44]Griep LM; Wolbers F; de Wagenaar B; ter Braak PM; Weksler BB; Romero IA; Couraud PO; Vermes I; van der Meer AD; van den Berg A, BBB on chip: microfluidic platform to mechanically and biochemically modulate blood-brain barrier function. Biomed Microdevices 2013, 15 (1), 145– 50. 22955726
  41. [45]Modarres HP; Janmaleki M; Novin M; Saliba J; El-Hajj F; RezayatiCharan M; Seyfoori A; Sadabadi H; Vandal M; Nguyen MD; Hasan A; Sanati-Nezhad A, In vitro models and systems for evaluating the dynamics of drug delivery to the healthy and diseased brain. J Control Release 2018, 273, 108– 130. 29378233
  42. [46]Bhalerao A; Sivandzade F; Archie SR; Chowdhury EA; Noorani B; Cucullo L, In vitro modeling of the neurovascular unit: advances in the field. Fluids Barriers CNS 2020, 17 (1), 22. 32178700 PMC7077137
  43. [47]Park TE; Mustafaoglu N; Herland A; Hasselkus R; Mannix R; FitzGerald EA; Prantil-Baun R; Watters A; Henry O; Benz M; Sanchez H; McCrea HJ; Goumnerova LC; Song HW; Palecek SP; Shusta E; Ingber DE, Hypoxia-enhanced Blood-Brain Barrier Chip recapitulates human barrier function and shuttling of drugs and antibodies. Nat Commun 2019, 10 (1), 2621. 31197168 PMC6565686
  44. [48]Shi L; Zeng M; Sun Y; Fu BM, Quantification of blood-brain barrier solute permeability and brain transport by multiphoton microscopy. J Biomech Eng 2014, 136 (3), 031005. 24193698
  45. [49]Shayan G; Choi YS; Shusta EV; Shuler ML; Lee KH, Murine in vitro model of the blood-brain barrier for evaluating drug transport. Eur J Pharm Sci 2011, 42 (1–2), 148– 55. 21078386
  46. [50]Silwedel C; Forster C, Differential susceptibility of cerebral and cerebellar murine brain microvascular endothelial cells to loss of barrier properties in response to inflammatory stimuli. J Neuroimmunol 2006, 179 (1–2), 37– 45. 16884785
  47. [51]Abbott NJ; Dolman DE; Drndarski S; Fredriksson SM, An improved in vitro blood-brain barrier model: rat brain endothelial cells co-cultured with astrocytes. In Methods in Molecular Biology (Methods and Protocols), 2011/12/07 ed.; Milner R, Ed. Humana Press: 2012; Vol. 814, pp 415– 30. 22144323
  48. [52]Hayashi K; Nakao S; Nakaoke R; Nakagawa S; Kitagawa N; Niwa M, Effects of hypoxia on endothelial/pericytic co-culture model of the blood-brain barrier. Regul Pept 2004, 123 (1–3), 77– 83. 15518896
  49. [53]Thomsen LB; Burkhart A; Moos T, A Triple Culture Model of the Blood-Brain Barrier Using Porcine Brain Endothelial cells, Astrocytes and Pericytes. PLoS One 2015, 10 (8), e0134765. 26241648 PMC4524625
  50. [54]Hatherell K; Couraud PO; Romero IA; Weksler B; Pilkington GJ, Development of a three-dimensional, all-human in vitro model of the blood-brain barrier using mono-, co-, and tri-cultivation Transwell models. J Neurosci Methods 2011, 199 (2), 223– 9. 21609734
  51. [55]Cucullo L; McAllister MS; Kight K; Krizanac-Bengez L; Marroni M; Mayberg MR; Stanness KA; Janigro D, A new dynamic in vitro model for the multidimensional study of astrocyte-endothelial cell interactions at the blood-brain barrier. Brain Res 2002, 951 (2), 243– 54. 12270503
  52. [56]Cucullo L; Hossain M; Tierney W; Janigro D, A new dynamic in vitro modular capillaries-venules modular system: cerebrovascular physiology in a box. BMC Neurosci 2013, 14, 18. 23388041 PMC3598202
  53. [57]Cucullo L; Hossain M; Puvenna V; Marchi N; Janigro D, The role of shear stress in Blood-Brain Barrier endothelial physiology. BMC Neurosci 2011, 12, 40. 21569296 PMC3103473
  54. [58]Booth R; Kim H, Characterization of a microfluidic in vitro model of the blood-brain barrier (muBBB). Lab Chip 2012, 12 (10), 1784– 92. 22422217
  55. [59]Wang YI; Abaci HE; Shuler ML, Microfluidic blood-brain barrier model provides in vivo-like barrier properties for drug permeability screening. Biotechnol Bioeng 2017, 114 (1), 184– 194. 27399645 PMC6650146
  56. [60]Brown JA; Pensabene V; Markov DA; Allwardt V; Neely MD; Shi M; Britt CM; Hoilett OS; Yang Q; Brewer BM; Samson PC; McCawley LJ; May JM; Webb DJ; Li D; Bowman AB; Reiserer RS; Wikswo JP, Recreating blood-brain barrier physiology and structure on chip: A novel neurovascular microfluidic bioreactor. Biomicrofluidics 2015, 9 (5), 054124. 26576206 PMC4627929
  57. [61]Herland A; van der Meer AD; FitzGerald EA; Park TE; Sleeboom JJ; Ingber DE, Distinct Contributions of Astrocytes and Pericytes to Neuroinflammation Identified in a 3D Human Blood-Brain Barrier on a Chip. PLoS One 2016, 11 (3), e0150360. 26930059 PMC4773137
  58. [62]Alcendor DJ, Human Vascular Pericytes and Cytomegalovirus Pathobiology. Int J Mol Sci 2019, 20 (6), 1– 12. PMC6471229 30909422
  59. [63]Shi J; Barakat M; Chen D; Chen L, Bicellular tight junction and wound healing. Int J Mol Sci 2018, 19 (12), 1– 20. PMC6321209 30518037

Cited by

Sorted by citations

Authors
Dong, X., & Shah, B.
Affiliations
University of North Texas Health Science Center (US)
Corresponding author
Dong, Xiaowei
ORCID
Venue
Current Drug Delivery
Publisher
Bentham Science Publishers Ltd.
Published
2022-12
Volume
19
Issue
10
Pages
1034-1046
ISSN
1567-2018 (ISSN-L)
DOI
10.2174/1567201819666220303102614
PMC
PMC9440157
OpenAlex
W4214861991
Cited by
42
References
4
Citations by year
2026: 8 · 2025: 12 · 2024: 18 · 2023: 4
FWCI
2.98
Type
journal article
Language
en
Open access
Locations
  • Current Drug Delivery (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
Funders
NIH, National Institutes of Health (1R35GM138225-01)
Topics
  • Life Sciences › Neuroscience › Neurology › Barrier Structure and Function Studies
  • Health Sciences › Medicine › Oncology › Drug Transport and Resistance Mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
Keywords
blood-brain barrier, in vitro BBB model, brain drug delivery, 3D model, human stem cells, blood-brain barrier permeability
MeSH terms
Biological Transport, Blood-Brain Barrier, Brain, Humans, Coculture Techniques, Endothelial Cells

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