Molecular Profiling of the Invasive Tumor Microenvironment in a 3-Dimensional Model of Colorectal Cancer Cells and <em>Ex vivo</em> Fibroblasts

Marc D. Bullock; Max Mellone; Karen M. Pickard; Abdulkadir Emre Sayan; Richard Mitter; John N. Primrose; Graham K. Packham; Gareth Thomas; Alexander H. Mirnezami

doi:10.3791/51475

Molecular Profiling von der invasiven Tumor-Microenvironment in einem 3-Dimensional Model of Colo...

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Molecular Profiling of the Invasive Tumor Microenvironment in a 3-Dimensional Model of Colorectal Cancer Cells and Ex vivo Fibroblasts

Molecular Profiling von der invasiven Tumor-Microenvironment in einem 3-Dimensional Model of Colorectal Cancer Cells und Ex-vivo- Fibroblasten

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10:33 min

April 29, 2014

DOI: 10.3791/51475-v

Marc D. Bullock^1,2, Max Mellone¹, Karen M. Pickard¹, Abdulkadir Emre Sayan¹, Richard Mitter³, John N. Primrose², Graham K. Packham¹, Gareth Thomas¹, Alexander H. Mirnezami^1,2

¹Cancer Sciences Unit,University of Southampton School of Medicine, ²University Surgical Unit,University of Southampton School of Medicine, ³Bioinformatics Unit,London Research Institute, Cancer Research UK

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Overview

This study presents a novel approach to characterize the biology at the interface between malignant epithelial cells and cancer-associated stromal cells in colorectal cancer models. By utilizing laser microdissection, researchers can dissect and analyze the distinct cellular environments within tumor microenvironments.

Key Study Components

Area of Science

Neuroscience
Oncology
Cell Biology

Background

Colorectal cancer involves complex interactions between tumor cells and the surrounding stroma.
Understanding these interactions is crucial for developing targeted therapies.
Laser microdissection allows for precise isolation of specific cell types within tumors.
This methodology can enhance molecular profiling efforts.

Purpose of Study

To characterize the biology at the invasive front of colorectal tumors.
To differentiate between invading and non-invading tumor cells.
To analyze the role of cancer-associated stromal cells in tumor progression.

Methods Used

Establishment of primary fibroblast cultures from colonic implants.
Construction of organotypic co-culture models with fibroblasts and colorectal cancer cells.
Fixation and sectioning of organotypic models for analysis.
Laser microdissection to separate tumor and stromal cells.

Main Results

Molecular profiling reveals distinct biological characteristics of tumor and stromal cells.
Identified differences in gene expression between invading and non-invading cells.
Insights into the tumor microenvironment's role in cancer progression.
Potential for developing targeted therapeutic strategies based on these findings.

Conclusions

This approach provides a manipulatable model for studying colorectal cancer biology.
Laser microdissection is a valuable tool for isolating specific cell populations.
Understanding the tumor-stroma interface can inform future cancer therapies.

Frequently Asked Questions

What is laser microdissection?

Laser microdissection is a technique used to isolate specific cells from a tissue sample using a laser.

Why is it important to study the tumor microenvironment?

The tumor microenvironment plays a critical role in cancer progression and response to therapy.

How do organotypic co-culture models work?

These models simulate the interactions between different cell types in a controlled environment, mimicking in vivo conditions.

What are the potential applications of this research?

Findings could lead to new therapeutic strategies targeting the tumor-stroma interactions in colorectal cancer.

What are cancer-associated stromal cells?

These are non-cancerous cells in the tumor microenvironment that can influence tumor growth and behavior.

What is the significance of profiling invading vs. non-invading cells?

Profiling these cells helps to understand the mechanisms of cancer invasion and metastasis.

Molecular Profiling von Laser mikrodissektiert Zellen und Stroma aus synthetischen, Tissue-Engineering-Darmkrebs-Modelle stellt einen neuartigen Ansatz zur manipulierbaren und an der Schnittstelle zwischen Tumorzellen zu charakterisieren und zu sezieren die unverwechselbare Biologie an der invasiven Front und Krebs Stromazellen.

Das übergeordnete Ziel dieses Verfahrens ist es, die charakteristische Biologie an der Schnittstelle zwischen malignen Epithelzellen an der invasiven Tumorfront und krebsassoziierten Stromazellen zu charakterisieren. Dies wird erreicht, indem zunächst primäre Fibroblastenkulturen aus Dickdarmimplantaten etabliert werden. Der zweite Schritt besteht darin, organotypische Co-Kulturmodelle unter Verwendung von primären Kolonfibroblasten und kultivierten Darmkrebszellen zu konstruieren.

Als nächstes werden organotypische Modelle in Formin fixiert, auf membrankaschierte Objektträger geschnitten und mit krestalem Violett gefärbt. Der letzte Schritt ist die Lasermikrodissektion, um eindringende Tumorzellen von nicht eindringenden Tumorzellen und krebsassoziierten Stromazellen zu trennen. Letztendlich können molekulare Profiling-Techniken verwendet werden, um die Biologie von eindringenden und nicht eindringenden Tumorzellen und Stromazellen in der umgebenden Tumormikroumgebung zu charakterisieren.

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