简介：
Overview
This article describes a procedure for establishing polarized liquid-covered cultures of Calu-3 cells using a Transwell culture system. The method emphasizes the advantages of using Calu-3 cells over normal human bronchial epithelial cells for rapid development into polarized cultures.
Key Study Components
Area of Science
- Cell culture techniques
- Polarization of epithelial cells
- Transwell culture systems
Background
- Calu-3 cells are a model for studying airway epithelial function.
- Polarized cultures are essential for understanding epithelial barrier properties.
- The Transwell system allows for the separation of apical and basolateral environments.
- Monitoring trans epithelial electrical resistance (TEER) is crucial for assessing polarization.
Purpose of Study
- To establish a reliable method for growing polarized Calu-3 cell cultures.
- To demonstrate the effectiveness of the Transwell system in cell culture.
- To provide a protocol for evaluating cell polarization using TEER and sodium fluorescein assays.
Methods Used
- Preparation of Transwell plates and cell suspensions.
- Seeding of Calu-3 cells into the apical compartments.
- Medium replacement on a specific cycle to promote polarization.
- Measurement of TEER and sodium fluorescein diffusion to assess polarization.
Main Results
- Successful establishment of polarized Calu-3 cultures.
- TEER measurements indicated complete polarization.
- Sodium fluorescein assays confirmed barrier integrity.
- The method proved to be efficient and reproducible.
Conclusions
- The Transwell culture system is effective for growing polarized Calu-3 cells.
- TEER and sodium fluorescein assays are reliable methods for evaluating cell polarization.
- This protocol can be utilized for further studies on airway epithelial function.
What are Calu-3 cells?
Calu-3 cells are a human bronchial epithelial cell line used as a model for studying airway epithelial function.
Why use a Transwell culture system?
The Transwell system allows for the separation of apical and basolateral environments, which is essential for studying polarized epithelial cells.
How is cell polarization assessed?
Cell polarization is assessed using trans epithelial electrical resistance (TEER) measurements and sodium fluorescein diffusion assays.
What is the significance of TEER?
TEER measurements provide insight into the integrity and barrier function of epithelial cell layers.
How long does it take to establish polarized cultures?
It typically takes two to three weeks to establish fully polarized cultures of Calu-3 cells.
What are the advantages of using Calu-3 cells?
Calu-3 cells are more accessible and develop more rapidly into polarized cultures compared to normal human bronchial epithelial cells.
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