<em>In vitro</em> Cell Culture Model for Toxic Inhaled Chemical Testing

Shama Ahmad; Aftab Ahmad; Keith B. Neeves; Tara Hendry-Hofer; Joan E. Loader; Carl W. White; Livia Veress

doi:10.3791/51539

In vitro Cell Culture Model for Toxic Inhaled Chemical Testing

JoVE Journal
Bioengineering

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JoVE Journal Bioengineering

In vitro Cell Culture Model for Toxic Inhaled Chemical Testing

In vitro Cell Culture Model for Toxic Inhaled Chemical Testing

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05:44 min

May 8, 2014

DOI: 10.3791/51539-v

Shama Ahmad¹, Aftab Ahmad¹, Keith B. Neeves², Tara Hendry-Hofer¹, Joan E. Loader¹, Carl W. White¹, Livia Veress¹

¹Pediatric Airway Research Center, Department of Pediatrics,University of Colorado, ²Department of Chemical and Biological Engineering,Colorado School of Mines

Overview

This protocol demonstrates the exposure method of cell cultures to inhaled toxic chemicals, specifically chlorine. It highlights the effects of chlorine exposure on airway epithelial cells and cardiomyocytes, providing a model for studying toxic gas interactions with vital organs.

Key Study Components

Area of Science

Neuroscience
Toxicology
Cell Biology

Background

Inhaled toxic chemicals can significantly affect lung and heart function.
Air-liquid interface cultures mimic human airway exposure to gases.
In vitro models allow for mechanistic studies without using whole animals.
Caspase activity and electrical resistance are key indicators of cell health.

Purpose of Study

To evaluate the toxicity of chlorine gas on cardiomyocytes and airway epithelial cells.
To model human exposure to toxic gases in a controlled environment.
To measure apoptotic cell death and membrane disruption in cell cultures.

Methods Used

Culturing rat cardiomyocytes and human airway epithelial cells.
Exposing cell cultures to chlorine gas.
Measuring caspase three and seven release.
Assessing trans epithelial electrical resistance.

Main Results

Chlorine exposure leads to significant cell death in both cardiomyocytes and airway epithelial cells.
Increased caspase activity indicates apoptosis in cardiomyocytes.
Disruption of membrane integrity observed in airway epithelial cells.
Electrical resistance measurements correlate with cell viability.

Conclusions

The study provides insights into the toxic effects of chlorine on lung and heart cells.
In vitro models are effective for studying the mechanisms of chemical toxicity.
Findings may inform the development of therapeutic strategies against chemical exposure.

Frequently Asked Questions

What is the significance of using in vitro models?

In vitro models allow researchers to study cellular responses to toxins without the ethical concerns associated with animal testing.

How does chlorine exposure affect cardiomyocytes?

Chlorine exposure can lead to increased apoptosis and cell death in cardiomyocytes, impacting heart function.

What measurements are taken to assess cell health?

Caspase activity and trans epithelial electrical resistance are measured to evaluate cell viability and membrane integrity.

Why is the air-liquid interface culture important?

It closely mimics the physiological conditions of the airway, providing a more accurate model for studying toxic gas exposure.

What are the potential applications of this research?

The findings could lead to the development of new therapeutic agents for treating injuries caused by toxic gas exposure.

Can this method be applied to other toxic substances?

Yes, the in vitro exposure system can be adapted to study various toxic chemicals and their effects on different cell types.

This protocol is designed to demonstrate exposure method of cell cultures to inhaled toxic chemicals. Exposure of differentiated air-liquid interface (ALI) cultures of airway epithelial cells provides a unique model of airway exposure to toxic gases such as chlorine. In this manuscript we describe effect of chlorine exposure on air-liquid interface cultures of epithelial cells and submerged culture of cardiomyocytes. In vitro exposure systems allow important mechanistic studies to evaluate pathways that could then be utilized to develop novel therapeutic agents.

The overall goal of the following experiment is to observe the effect of chlorine exposure in in vitro cell culture models of the lung, airway and heart. This is achieved by first culturing rat cardiomyocytes or human airway epithelial cells. The rat cardiomyocyte cultures will be exposed to chlorine gas to evaluate the chlorine toxicity against vital organs such as the heart, whereas the exposure of differentiated cultures of human airway, epithelial cells to chlorine allows the further modeling of human exposures to toxic gases.

Ultimately, caspase three seven release and trans epithelial electrical resistance can be measured to determine the apoptotic cell death of the cardiomyocytes and the membrane disruption and cell death of the human airway epithelial cells respectively. The main advantage of this method is that it involves primary tissue cell cultures instead of whole animals, hence allowing a simple in vitro chemical testing method. Tip four, perform chlorine exposure experiments.

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