Journals / Kafkas Üniversitesi Veteriner Fakültesi Dergisi / 2018 / Cilt: 24 - Sayı: 3

In vitro Model of the Blood-brain Barrier Established by Co-culture of Primary Sheep Cerebral Microvascular Endothelial and Astrocyte Cells

Primer Koyun Serebral Mikrodamar Endotelyal ve Astrosit Hücrelerinin Birlikte Kültüre Edilmesiyle Oluşturulmuş In vitro Kan Beyin Bariyeri Modeli

Pages
379–386
Publication Date
2022-10-18
DOI
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AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

Results showed that our model may be avaluable tool in the study of the mechanisms of brain invasion mediated by internalins of Listeria monocytogenes.

📌 3 Key Takeaways

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Objective & Hypothesis:Blood-brain barrier (BBB) characteristics are induced and maintained by cross-talk between brain microvessel endothelial cells andneighbouring elements of the neurovascular unit.
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Key Findings & Data:Here, we have developed and characterized an in vitro model of the blood-brain barrier by seeding primarybrain microvascular endothelial cells (BMECs) and astrocyte cells (ACs) from sheep on a polyester Transwell cell culture membrane with0.4-μm pores, and conducted transepithelial electrical resistance measurements (TEER), leakage tests and assays for the permeability ofhorseradish peroxidase (HRP).
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Conclusion & Impact:Results showed that our model may be avaluable tool in the study of the mechanisms of brain invasion mediated by internalins of Listeria monocytogenes.
🔬Design:Modeling & Simulation
📍Scope:Comprehensive Sample
⚙️Variables:Domain Parameters

Abstract

Blood-brain barrier (BBB) characteristics are induced and maintained by cross-talk between brain microvessel endothelial cells andneighbouring elements of the neurovascular unit. It plays a primary role in a selective diffusion barrier at the level of the cerebralmicrovascular endothelium. Here, we have developed and characterized an in vitro model of the blood-brain barrier by seeding primarybrain microvascular endothelial cells (BMECs) and astrocyte cells (ACs) from sheep on a polyester Transwell cell culture membrane with0.4-μm pores, and conducted transepithelial electrical resistance measurements (TEER), leakage tests and assays for the permeability ofhorseradish peroxidase (HRP). The results of TEER and 4-h leakage assay showed that the new syngeneic BBB model had a characteristicof BBB in vivo. The HRP permeability test showed that the BBB model had good barrier function. Results showed that our model may be avaluable tool in the study of the mechanisms of brain invasion mediated by internalins of Listeria monocytogenes.

Özet

Kan beyin bariyeri beyin mikrodamar endotelyal hücreleri ve nörovasküler birimin komşu elementleri tarafından oluşturulur ve devamlılığı sağlanır. Kan beyin bariyeri serebral mikrovasküler endotel seviyesinde seçici difüzyon bariyeri olarak önemli bir rol oynar. Bu çalışmada 0.4- μm gözeneğe sahip polyester Transwell hücre kültürü membranına koyun primer beyin microdamar endotel hücreleri ve astrosit hücrelerinin ekimiyle in vitro kan beyin bariyeri oluşturuldu. Karakterizasyonu transepitelyal elektrik direnç ölçümleri, sızıntı testleri ve horseradish peroksidaz geçirgenliği testleri ile gerçekleştirildi. Transepitelyal elektrik direnç ölçümleri ve 4-h sızıntı testleri sonuçları yeni oluşturulan singeneik kan beyin bariyeri modelinin in vivo kan beyin bariyeri özelliklerine sahip olduğunu gösterdi. Horseradish peroksidaz geçirgenliği testi yeni oluşturulan kan beyin bariyerinin iyi bariyer fonksiyonlarına sahip olduğunu gösterdi. Elde edilen sonuçlar oluşturulan Listeria monocytogenes internalinleri tarafından oluşturulan beyin invazyonunun mekanizmasının çalışılmasında bu modelin yararlı olabileceğini göstermektedir.

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Cite This Article

REN, J., YANG, M., YAN, G., JIANG, J., WANG, P. (2022). In vitro Model of the Blood-brain Barrier Established by Co-culture of Primary Sheep Cerebral Microvascular Endothelial and Astrocyte Cells. Kafkas Üniversitesi Veteriner Fakültesi Dergisi, Cilt: 24 - Sayı: 3, 379–386. https://ajindex.com/articles/569820

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References (4)

  1. Tian X, Brookes O, Battaglia G: Pericytes from mesenchymal stem cells as a model for the blood-brain barrier. Sci Rep, 7, 39676, 2017. DOI: 10.1038/srep39676 Scism JL, Laska DA, Horn JW, Gimple JL, Pratt SE, Shepard RL, Dantzig AH, Wrighton SA: Evaluation of an in vitro, co-culture model for the blood-brain barrier: Comparison of human umbilical vein endothelial cells (ECV304) and rat glioma cells (C6) from two commercial sources. In Vitro Cell Dev Biol Anim, 35 (10): 580-592, 1999. DOI: 10.1007/s11626-999- 0096-3
  2. Schroeter ML, Müller S, Lindenau J, Wiesner B, Hanisch UK, Wolf G, Blasig IE: Astrocytes induce manganese superoxide dismutase in brain capillary endothelial cells. Neuroreport, 12 (11): 2513-2517, 2001. DOI: 10.1097/00001756-200108080-00045
  3. 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, 12 (3): 215-222, 2001. DOI: 10.1016/S0928- 0987(00)00123-8
  4. Kaisar MA, Sajja RK, Prasad, S, Abhyankar VV, Liles T, Cucullo L: New experimental models of the blood-brain barrier for CNS drug discovery. Expert Opin Drug Discov, 12 (1): 89-103, 2017. DOI: 10.1080/17460441.2017.1253676 Rapôso C, Odorissi PAM, Oliveira ALR, Aoyama H, Ferreira CV, Verinaud L, Fontana K, Ruela-de-Sousa RR, da Cruz-Höfling MA: Effect of Phoneutria nigriventer, venom on the expression of junctional protein and P-gp efflux pump function in the blood–brain barrier. Neurochem Res, 37 (9): 1967-1981, 2012. DOI: 10.1007/s11064-012-0817-y Kim KS: Microbial translocation of the blood-brain barrier. Int J Parasitol, 36 (5): 607-614, 2006. DOI: 10.1016/j.ijpara.2006.01.013 Hanada S, Fujioka K, Inoue Y, Kanaya F, Manome Y, Yamamoto K: Cell-based in vitro blood-brain barrier model can rapidly evaluate nanoparticles’ brain permeability in association with particle size and surface modification. Int J Mol Sci, 15 (2): 1812-1825, 2014. DOI: 10.3390/ ijms15021812 Wuest DM, Lee KH: Optimization of endothelial cell growth in a murine in vitro blood-brain barrier model. Biotechnol J, 7 (3): 409-417, 2012. DOI: 10.1002/biot.201100189 Burek M, Salvador E, Förster CY: Generation of an immortalized murine brain microvascular endothelial cell line as an in vitro blood brain barrier model. J Vis Exp, 66, e4022, 2012. DOI: 10.3791/4022 Steiner O, Coisne C, Engelhardt B, L