Optimization of the Wound Scratch Assay to Detect Changes in Murine Mesenchymal Stromal Cell Migration After Damage by Soluble Cigarette Smoke Extract

Nicholas Cormier; Alexander Yeo; Elizabeth Fiorentino; Julia Paxson

doi:10.3791/53414

Optimization of the Wound Scratch Assay to Detect Changes in Murine Mesenchymal Stromal Cell Migr...

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Developmental Biology

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JoVE Journal Developmental Biology

Optimization of the Wound Scratch Assay to Detect Changes in Murine Mesenchymal Stromal Cell Migration After Damage by Soluble Cigarette Smoke Extract

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08:40 min

December 3, 2015

DOI: 10.3791/53414-v

Nicholas Cormier*¹, Alexander Yeo*¹, Elizabeth Fiorentino¹, Julia Paxson¹

¹Department of Biology,College of the Holy Cross

Overview

This article presents a migration assay designed to quantify changes in the migratory capacity of lung mesenchymal stromal cells (MSCs) after exposure to soluble cigarette smoke extract. The method is optimized for better quantitative measurements while remaining economical and customizable.

Key Study Components

Area of Science

Cell migration
Lung regenerative biology
Mesenchymal stromal cells

Background

Mesenchymal stromal cells play a crucial role in tissue repair.
Quantifying migratory capacity is essential for understanding cell behavior post-damage.
Existing methods may lack rigorous statistical analysis.
This study focuses on lung MSCs and their response to environmental stressors.

Purpose of Study

To detect changes in MSC migratory capacity after damage.
To improve understanding of lung progenitor cell migration.
To provide a reliable method for assessing cell migration in regenerative biology.

Methods Used

Scratch assay to quantify cell migration.
Use of soluble cigarette smoke extract to induce damage.
Growth of MSCs in complete media on 60 mm plates.
Statistical analysis to assess changes in migration.

Main Results

Demonstrated the impact of cigarette smoke extract on MSC migration.
Provided quantitative measurements of migratory capacity.
Showed that the scratch assay can effectively measure changes in cell behavior.
Highlighted the importance of confluence for accurate imaging results.

Conclusions

The assay is a valuable tool for studying MSC migration.
It can help answer critical questions in lung regenerative biology.
Future applications may lead to improved strategies for lung repair.

Frequently Asked Questions

What is the main goal of the migration assay?

To detect changes in the migratory capacity of lung mesenchymal stromal cells after damage.

How does the scratch assay work?

The scratch assay quantifies cell migration by creating a scratch in a cell monolayer and measuring the closure over time.

What is the significance of using soluble cigarette smoke extract?

It simulates environmental damage to study its effects on lung progenitor cell migration.

Why is confluence important in this assay?

Proper confluence ensures accurate identification of scratch boundaries for imaging analysis.

Can this method be customized?

Yes, the assay is designed to be easily adaptable for various experimental needs.

What are the advantages of this migration assay?

It provides better quantitative measurements and is economical to perform.

The capacity to migrate is a key function of many different cell types, including mesenchymal stromal cells (MSCs). However, quantifying alterations in migratory capacity after damage is challenging. This protocol describes an easily adaptable migration assay that uses rigorous statistics to quantify changes in MSC migratory capacity after damage.

The overall goal of this assay is to detect changes in the migratory capacity of lung mesenchymal stromal cells after damage with soluble cigarette smoke extract. This method can be used to answer key questions in the field of lung regenerative biology, including how migratory capacity of lung progenitor cells can be improved to aid in lung repair. The main advantage of this technique is that it's been optimized to provide better quantitative measurements for cell migration while still remaining economical to perform and easy to customize us.

In this demonstration, the scratch assay will be used to quantify the migratory capacity of murine lung mesenchymal stromal cells, or MSCs. After exposure to soluble cigarette smoke extract, the MSCs are grown in complete media on 60 millimeter plates and damaged as described in the protocol text. An appropriate confluence is important for the scratch assay because it ensures that the imaging software can correctly identify the boundaries of the scratch.

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