Shear Stress and Barrier Function
Blood vessels in the central nervous system (CNS) and peripheral nervous system (PNS) are comprised of endothelial cells, which are subjected to constant fluid flow. This fluid flow generates shear stress, which significantly impacts the morphology, function, and behavior of these cells.
For example, shear stress can induce the production of reactive oxygen species (ROS) and the release of inflammatory cytokines in brain endothelial cells. This can lead to oxidative stress and inflammation, which have been implicated in the pathogenesis of neurological disorders such as stroke and Alzheimer's disease.
The blood-brain-barrier (BBB) is a highly selective and dynamic barrier that separates the brain from circulating blood. It plays a critical role in maintaining brain homeostasis and protecting the brain from harmful substances. Cell culture under flow assays can be used to model the BBB in vitro, which allows researchers to study the mechanisms of BBB function and dysfunction and to screen drugs that can penetrate the BBB. In addition, the function and the effects of drugs and toxins on the barrier can be investigated with Electric Cell-substrate Impedance Sensing (ECIS). In this assay, changes in impedance of a cell monolayer are measured in response to electrical stimulation.
ibidi Solutions for Shear Stress Assays and Barrier Function
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The ibidi Pump System is ideal for long-term cell culture under flow with defined shear stress values and is compatible with all µ-Slides with Luer adapters. It simulates defined continuous and pulsatile laminar flow and oscillatory flow to study cells in a more physiological environment. It is optimal for rolling and adhesion assays, transmigration, and invasion studies. Also, cells, spheroids, and organoids can be perfused for optimal nutrition. |
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ibidi provides a variety of Channel Slides with different geometries. The µ-Slide I Luer family has one channel for standard flow experiments and rolling and adhesion assays. The µ-Slide VI has 6 channels and can be used for parallel flow assays. Both are available with the ibidi Polymer Coverslip and the ibidi Glass Coverslip, plus different heights and coatings. |
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The μ-Slide I Luer 3D can help create an endothelial barrier without the need of an artificial filter membrane. Endothelial cells can be seeded on a suitable gel matrix, such as Collagen Type I. After connecting the slide to a pump and applying defined shear stress, an in vivo-like endothelial barrier is created, which is useful for rolling and adhesion assays). |
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The impedance measurement system ECIS Model Z Theta allows for label-free, non-invasive cell-based assays to measure changes in morphology and cellular function (such as barrier function). It is available with a 16 Well Array Station for 2x ECIS Cultureware 8 well arrays and a 96 Well Array Station for ECIS Cultureware 96 well arrays. The ECIS Model Z Theta is compatible with ECIS Cultureware in an open 8 well slide format or 96 well multiwell format. Both formats are available with different array arrangements and geometries. |
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The ECIS system can be combined with the ECIS Flow Module for long-term cell culture under flow with defined shear stress values and is compatible with ECIS Flow Arrays. It allows for impedance measurements under flow. |
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The ECIS Flow Arrays can be used for endothelial cell and cell-cell interaction studies at defined flow rates. Flow Arrays with one channel are suitable for standard flow experiments and rolling and adhesion assays. The ECIS Flow Arrays 6 Channel µ-Slide has 6 channels and can be used for parallel flow assays. Both are available with different electrode numbers and arrangements. |
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Endothelial cells can be seeded on a suitable gel matrix, such as Collagen Type I (bovine or rat tail origin). Find more information in AN 26: Collagen I Gel for 3D Cell Culture. |
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The ibidi Stage Top Incubators provide physiological conditions for live cell imaging on every standard inverted microscope. They include CO2 and O2 control (e.g., for hypoxia experiments) as well as actively controlled humidity. They are available for single slides and dishes or for multiwell plates. |
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Selected References
Cultivation of Brain Microvascular Endothelial Cells (BMECs) Under Flow
To investigate the effect of shear stress on gerbil brain microvascular endothelial cells (BMECs), cells were seeded into the µ-Slide I 0.6 Luer and subjected to unidirectional flow with the ibidi Pump System.
Gao JQ, Wang P, Yan JW, Ba LN, Shi PL, Wu HM, Guan XY, Cao YG, Sun HL, Mao XY. Shear Stress Rescued the Neuronal Impairment Induced by Global Cerebral Ischemia Reperfusion via Activating PECAM-1-eNOS-NO Pathway. Front Cell Dev Biol. 2021 Jan 21;8:631286. doi: 10.3389/fcell.2020.631286.
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Cultivation of Astrocytes Under Flow
Astrocytes were incubated at physiological conditions using the ibidi Stage Top Incubator and exposed to shear stress using the ibidi Pump System.
Wakida NM, Cruz GMS, Pouladian P, Berns MW, Preece D. Fluid Shear Stress Enhances the Phagocytic Response of Astrocytes. Front Bioeng Biotechnol. 2020 Nov 11;8:596577. doi: 10.3389/fbioe.2020.596577.
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in vitro Blood Brain Barrier (BBB) Model
Assembly of a blood brain barrier (BBB) in vitro system using the ibidi Pump System to check nanoparticles (for treatment of glioblastoma multiforme) for their ability to cross the barrier.
Grillone A, Battaglini M, Moscato S, Mattii L, de Julián Fernández C, Scarpellini A, Giorgi M, Sinibaldi E, Ciofani G. Nutlin-loaded magnetic solid lipid nanoparticles for targeted glioblastoma treatment. Nanomedicine (Lond). 2019 Mar;14(6):727-752. doi: 10.2217/nnm-2018-0436.
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Cell Culture Under Flow of Brain Microvascular Endothelial Cells (BMECs)
Cultivation of BMECs under flow using the µ-Slide I 0.6 Luer and the ibidi Pump system to investigate the effect of shear stress.
Gao JQ, Wang P, Yan JW, Ba LN, Shi PL, Wu HM, Guan XY, Cao YG, Sun HL, Mao XY. Shear Stress Rescued the Neuronal Impairment Induced by Global Cerebral Ischemia Reperfusion via Activating PECAM-1-eNOS-NO Pathway. Front Cell Dev Biol. 2021 Jan 21;8:631286. doi: 10.3389/fcell.2020.631286.
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Shear Stress Model to Study BBB Permeability
Establishing a shear stress model using human cerebral microvascular endothelial cells (hCMEC) and the ibidi Pump System to investigate drug permeability of the BBB.
Choublier N, Müller Y, Gomez Baisac L, Laedermann J, de Rham C, Declèves X, Roux A. Blood–Brain Barrier Dynamic Device with Uniform Shear Stress Distribution for Microscopy and Permeability Measurements. Applied Sciences 2021 Jun 16;11(12), 5584. doi: 10.3390/app11125584.
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Impedance Measurements to Investigate BBB Function in Alzheimer’s Disease Development
Investigating the effects of different Alzheimer’s disease amyloid β heterogeneous fragments on BBB function using the ECIS Model Zθ.
Parodi-Rullán R, Ghiso J, Cabrera E, Rostagno A, Fossati S. Alzheimer's amyloid β heterogeneous species differentially affect brain endothelial cell viability, blood-brain barrier integrity, and angiogenesis. Aging Cell. 2020 Nov;19(11):e13258. doi: 10.1111/acel.13258.
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Read on and learn more about Neuronal Function and Activity, Vascularization / Angiogenesis, and Neural Migration and Chemotaxis.