bend5 cell line  (Weksler)

Journal: International Journal of Molecular Sciences

Article Title: The Role of Cytoskeletal Proteins in the Formation of a Functional In Vitro Blood-Brain Barrier Model

doi: 10.3390/ijms23020742

Figure Lengend Snippet: HR-SEM micrographs representing BEC bEnd5 NTs that are subdivided into two types: ( A ) NV-induced TUNTs indicated by the purple circles, scale bar 1000 nm; ( B ) TUNT formation and extension across the PC space between adjacent BECs, upon the fusion of multiple NVs, scale bar = 300 nm; ( C ) TENTs formed by the BEC membrane leading edges as indicated by the yellow circles, scale bar = 1000 nm; ( D ) A magnified TENT extension across the PC space between two adjacent BECs forming an occluded tent-like covering, scale bar = 300 nm.

Article Snippet: The bEnd5 cell line was purchased from the European Collection of Authenticated Cell Cultures (ECACC) (Sigma-Aldrich, 96091930, St. Louis, MI, USA).

Techniques: Membrane

Journal: International Journal of Molecular Sciences

Article Title: The Role of Cytoskeletal Proteins in the Formation of a Functional In Vitro Blood-Brain Barrier Model

doi: 10.3390/ijms23020742

Figure Lengend Snippet: High-resolution scanning electron microscopy (HR-SEM) micrographical representation of the morphological deformation of cell-cell connections between adjacent BECs upon the chemical perturbation of actin polymerization utilizing a known depolymerizing agent, Cytochalasin D. ( A ) Indicates the numerous TENT interactions across the PC space between two bEnd5 cells. The micrograph clearly depicts the extensive formation of tethering-NTs which are naturally taut and project across the PC space; ( B ) A micrograph demonstrating the depolymerizing effect of Cytochalasin D on F-actin of bEnd5 cells where TENT formation is compromised forming thick, brittle-type NTs, as well as a distinct decrease in the TENT formation. An observable decrease in the number of NVs on the treated BEC membrane was seen, compared to the numerous NVs that were located on the control BEC membranes in ( A ). The yellow star represents cell 1, the blue star represents cell 2, and the paracellular space is denoted by PC .

Article Snippet: The bEnd5 cell line was purchased from the European Collection of Authenticated Cell Cultures (ECACC) (Sigma-Aldrich, 96091930, St. Louis, MI, USA).

Techniques: Electron Microscopy, Membrane, Control

Journal: International Journal of Molecular Sciences

Article Title: The Role of Cytoskeletal Proteins in the Formation of a Functional In Vitro Blood-Brain Barrier Model

doi: 10.3390/ijms23020742

Figure Lengend Snippet: HR-SEM micrographical representation of the morphological deformation of cell-cell connections between adjacent BECs upon Nocodazole-induced cytoarchitectural depolymerization. ( A ) The bEnd5 TENT formation with cells that were grown in standard culture medium. The yellow arrows show the normal nature of PC TENTs, reflecting their bearing and tautness; ( B ) The effect of nocodazole exposure on bEnd5 NT formation relative to the control. The yellow star represents cell 1, the blue star represents cell 2, and the paracellular space is denoted by PC . The red circles in ( B ) indicate NTs which have lost their tautness and demonstrate the recoiling of TENTs in the Nocodazole-treated bEnd5 cell cultures.

Article Snippet: The bEnd5 cell line was purchased from the European Collection of Authenticated Cell Cultures (ECACC) (Sigma-Aldrich, 96091930, St. Louis, MI, USA).

Techniques: Control

Journal: International Journal of Molecular Sciences

Article Title: The Role of Cytoskeletal Proteins in the Formation of a Functional In Vitro Blood-Brain Barrier Model

doi: 10.3390/ijms23020742

Figure Lengend Snippet: The effect of known depolymerizing agent Cytochalasin D on the physiological parameters of BECs. ( A ) The effect of 0.1–1 μM Cytochalasin D on the TEER across the confluent monolayers of bEnd5 BECs after 12–24 h exposure. The single asterisk * represents significant differences between the experiments and the control samples. The double asterisk (**) denotes statistical significance between the experimental samples and the vehicle control 2 (C2). The data are expressed as the mean ± SEM; ( B ) represents the effect of Cytochalasin D on bEnd5 cell numbers. The asterisks * denotes statistically significant differences between the experimental samples relative to control 1 (C1) and the vehicle control 2 (C2- with 0.1% DMSO); ( C ) Assessing the depolymerizing effect of Cytochalasin D in IF micrographs of Alexa Fluor 488 conjugated, monoclonal F-actin antibodies (Ab’), the yellow arrowheads indicate the regions of NT formation and the red arrowheads indicate the regions of depolymerization and/or intercellular gaps at the membrane leading edges between adjacent BECs, 12 h and 24 h. D (Scale bar = 50 μM) ( D ) Western blot analysis displays the effect of chemical perturbation of Cytochalasin D on F-actin expression. The asterisks * denotes statistically significant differences between the experimental samples and both control 1 and 2. The data are represented as the mean ± SEM ( n = 3). Statistical significance was determined at p < 0.05.

Article Snippet: The bEnd5 cell line was purchased from the European Collection of Authenticated Cell Cultures (ECACC) (Sigma-Aldrich, 96091930, St. Louis, MI, USA).

Techniques: Control, Membrane, Western Blot, Expressing

Journal: International Journal of Molecular Sciences

Article Title: The Role of Cytoskeletal Proteins in the Formation of a Functional In Vitro Blood-Brain Barrier Model

doi: 10.3390/ijms23020742

Figure Lengend Snippet: The effect of Nocodazole on the selected physiological parameters of bEnd5 cells. ( A ) The decreasing TEER across confluent bEnd5 cell monolayers upon treatment with 0.25–2 μM Nocodazole. The single asterisk * represents significant differences between the experiments and the untreated (control) samples. The data are expressed as the mean ± SEM; ( B ) Represents the effect of Nocodazole on bEnd5 cell numbers. The asterisk * denotes statistically significant differences between the experimental samples relative to the untreated conditions (C1) and the vehicle control 2 (C2-with 0.1% DMSO); ( C ) IF micrographs of Alexa Fluor 488 conjugated, monoclonal α-tubulin Ab’ after 24–48 h Nocodazole treatment for 24 h (Scale bar = 50 μM) and 48 h (Scale bar = 20 μM). The yellow arrowheads indicate regions of intercellular NT formation and the red arrowheads indicate regions of depolymerization at the membrane leading edges between adjacent BECs; ( D ) The depolymerizing effect of Nocodazole on α-tubulin at 24 h and 48 h observed in the Western blot analysis, which showed significant suppression in protein expression relative to the controls for 24–48 h. The asterisks * denotes statistically significant differences between the experimental samples and both control 1 and 2. The data are represented as the mean ± SEM ( n = 3). Statistical significance was determined at p -value < 0.05.

Article Snippet: The bEnd5 cell line was purchased from the European Collection of Authenticated Cell Cultures (ECACC) (Sigma-Aldrich, 96091930, St. Louis, MI, USA).

Techniques: Control, Membrane, Western Blot, Expressing