Barrier Functional Integrity Recording on bEnd.3 Vascular Endothelial Cells <em>via</em> Transendothelial Electrical Resistance Detection

Fuhan Fan; Hong Jiang; Ya Hou; Yi Zhang; Qipeng Zhao; Yong Zeng; Xianli Meng; Xiaobo Wang

doi:10.3791/65938

Registro de la integridad funcional de la barrera en las células endoteliales vasculares...

JoVE Journal
Neuroscience

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Neuroscience

Barrier Functional Integrity Recording on bEnd.3 Vascular Endothelial Cells via Transendothelial Electrical Resistance Detection

Registro de la integridad funcional de la barrera en las células endoteliales vasculares bEnd.3 mediante detección de resistencia eléctrica transendotelial

Full Text

4,767 Views

09:03 min

September 29, 2023

DOI: 10.3791/65938-v

Fuhan Fan¹, Hong Jiang¹, Ya Hou¹, Yi Zhang^1,2,3, Qipeng Zhao⁴, Yong Zeng^1,2, Xianli Meng^1,2,5, Xiaobo Wang²

¹School of Pharmacy/School of Modern Chinese Medicine Industry,Chengdu University of Traditional Chinese Medicine, ²Meishan Hospital of Chengdu University of Traditional Chinese Medicine, ³School of Ethnic Medicine,Chengdu University of Traditional Chinese Medicine, ⁴School of Pharmacy,Ningxia Medical University, ⁵Innovative Institute of Chinese Medicine and Pharmacy/Academy for Interdiscipline,Chengdu University of Traditional Chinese Medicine

Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This study presents a reliable in vitro model of the blood-brain barrier (BBB) using mouse cerebral vascular endothelial cells (bEnd.3). The model assesses transmembrane electrical resistance (TER) to evaluate the integrity of the BBB, facilitating drug screening for central nervous system treatments.

Key Study Components

Area of Science

Cell Biology
Neuroscience
Pharmacology

Background

The blood-brain barrier is crucial for maintaining homeostasis in the central nervous system.
Traditional in vitro models are less effective for drug testing due to limitations in cellular complexity.
This study addresses those limitations by presenting a simpler, non-invasive method.

Purpose of Study

To improve the assessment of BBB integrity through TER measurement.
To facilitate the screening of drugs targeting central nervous system disorders.
To verify the relationship between TER values and tight junction protein expression.

Methods Used

The study utilized bEnd.3 cell culture, which was seeded and cultured in a 96-well plate format.
Key interventions included the addition of cobalt chloride to assess cellular viability and barrier integrity.
TEER values were recorded over time to determine the formation and stability of the BBB model.
Statistical analysis was employed to compare control and treated groups.

Main Results

Results indicated a significant reduction in TEER values upon treatment with cobalt chloride, indicating compromised BBB integrity.
The study showed the potential of using TER as a reliable method for assessing drug effects on the BBB.
Stability of the cell barrier was reached by day five, underscoring the method's effectiveness.

Conclusions

This study demonstrates a simplified and effective approach to assess BBB integrity in drug development.
Results could enhance understanding of therapeutic interventions for central nervous system diseases.
The methods outlined may be adapted for broader applications in pharmacological research.

Frequently Asked Questions

What are the advantages of using the bEnd.3 cell line?

The bEnd.3 cell line is an established model for studying the blood-brain barrier, providing relevant insights into endothelial function and drug permeability.

How is the cobalt chloride treatment implemented?

Cobalt chloride is added to the cell cultures to investigate its effects on cell viability and barrier function, specifically assessing changes in TER.

What measures are taken to ensure accurate TEER readings?

Calibration of the measurement instrument and proper handling of electrodes are critical for obtaining reliable TEER measurements before and during the experiments.

What types of data can be obtained from this method?

The method provides data on electrical resistance, supporting assessments of cell viability and barrier integrity essential for drug development.

How can this model be adapted for other studies?

This BBB model could be modified to test various compounds or treatments, enhancing its utility for pharmacological and toxicological research.

What are the limitations of this study?

While the model is reliable, it may not fully replicate in vivo conditions, signaling the need for complementary studies using more complex biological models.

Este protocolo describe un modelo in vitro fiable y eficiente de la barrera hematoencefálica. El método utiliza células endoteliales vasculares cerebrales de ratón bEnd.3 y mide la resistencia eléctrica transmembrana.

Los estudios actuales suelen utilizar pruebas de permeabilidad y la expresión de proteínas asociadas para verificar la precisión de la detección de TEER. El modelo BBB de células monocapa en virtud de ella es menos eficaz para el cribado de fármacos, y el uso de más células dificulta la construcción del modelo. El protocolo de este estudio es relativamente sencillo, ya que el procedimiento no invasivo permite utilizar las muestras celulares en otros experimentos tras la detección de TEER.

La detección de TEER para evaluar la permeabilidad de los modelos celulares puede acelerar el cribado de fármacos para el tratamiento y la prevención del sistema nervioso central con el fin de aumentar la tasa de éxito del desarrollo de nuevos fármacos. En el futuro, detectaremos la expresión de la proteína de unión estrecha en las células del compartimento superior de la pared torácica para verificar que los resultados de la prueba TEER puedan representar eficazmente la integridad de la función de barrera de la BHE.

View the full transcript and gain access to thousands of scientific videos

Explore More Videos

Registro de la Integridad Funcional de la Barrera Células Endoteliales Vasculares BEnd.3 Detección de Resistencia Eléctrica Transendotelial Barrera Hematoencefálica Células Endoteliales Microvasculares Astrocitos Pericitos Modelos In Vitro Trastornos Neurológicos Tratamientos Farmacológicos Modelo de Células BBB Monocapa Cámara Superior Placa de 24 Pocillos Función de Barrera Celular Voltímetro de Células Epiteliales Resistencia Eléctrica Transmembrana Células Hipóxicas Inducidas por CoCl2 Enfermedades del Sistema Nervioso Central