Intramucosal Inoculation of Squamous Cell Carcinoma Cells in Mice for Tumor Immune Profiling and Treatment Response Assessment

Ayman  J. Oweida; Shilpa Bhatia; Benjamin Van Court; Laurel Darragh; Natalie Serkova; Sana  D. Karam

doi:10.3791/59195

Intramucosal Impfung des Plattenepithelkarzinom Zellen in Mäusen für Tumor immun Profilierung und...

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Cancer Research

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JoVE Journal Cancer Research

Intramucosal Inoculation of Squamous Cell Carcinoma Cells in Mice for Tumor Immune Profiling and Treatment Response Assessment

Intramucosal Impfung des Plattenepithelkarzinom Zellen in Mäusen für Tumor immun Profilierung und Behandlung Antwort Bewertung

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07:29 min

April 22, 2019

DOI: 10.3791/59195-v

Ayman J. Oweida¹, Shilpa Bhatia¹, Benjamin Van Court¹, Laurel Darragh¹, Natalie Serkova^1,2,3, Sana D. Karam¹

¹Department of Radiation Oncology,University of Colorado Denver - Anschutz Medical Campus, ²Department of Anesthesiology,University of Colorado Denver - Anschutz Medical Campus, ³Division of Radiology,University of Colorado Denver - Anschutz Medical Campus

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Overview

This article presents a reproducible method for developing an orthotopic murine model of head and neck squamous cell carcinoma. The model exhibits clinically relevant histopathological features and symptoms similar to those observed in human patients.

Key Study Components

Area of Science

Oncology
Neuroscience
Pathology

Background

Head and neck cancers are increasingly prevalent.
Understanding the tumor microenvironment is limited.
Mechanisms of treatment resistance in head and neck tumors are not well understood.
This model recapitulates the native microenvironment of these tumors.

Purpose of Study

To develop a method for creating a murine model of head and neck squamous cell carcinoma.
To study the tumor microenvironment and treatment resistance.
To observe clinical manifestations similar to those in humans.

Methods Used

Cell culture techniques for tumor cell preparation.
Orthotopic implantation of tumor cells in mice.
Monitoring of tumor growth and clinical symptoms.
Histopathological analysis of tumor samples.

Main Results

Developed tumors show necrosis and poor differentiation.
Presence of nodal metastases and immune infiltration.
Clinical symptoms include dysphagia, jaw displacement, and weight loss.
The model effectively mimics human disease characteristics.

Conclusions

The orthotopic murine model is a valuable tool for studying head and neck squamous cell carcinoma.
This model can aid in understanding treatment resistance mechanisms.
It provides insights into the tumor microenvironment and clinical manifestations.

Frequently Asked Questions

What is the significance of this murine model?

It allows researchers to study head and neck squamous cell carcinoma in a setting that closely resembles human disease.

How does this model mimic human symptoms?

The model exhibits symptoms such as dysphagia and jaw displacement, which are clinically relevant to human patients.

What are the histopathological features observed?

Tumors show necrosis, poor differentiation, and immune infiltration, similar to human tumors.

What methods are used to analyze the tumors?

Histopathological analysis and monitoring of clinical symptoms are key methods used in this study.

Why is understanding the tumor microenvironment important?

It helps in identifying mechanisms of treatment resistance and developing more effective therapies.

What are the potential applications of this research?

The findings can inform future studies on treatment strategies and improve understanding of head and neck cancers.

Hier präsentieren wir eine reproduzierbare Methode für die Entwicklung einer orthotopen Mausmodell von Kopf und Hals Squamous Zelle Krebsgeschwür. Tumoren zeigen klinisch relevante histopathologischen Merkmale der Krankheit, einschließlich der Nekrose, schlechte Differenzierung und Knotenpunkte Metastasen immun Infiltration. Tumor-tragenden Mäusen Symptome klinisch relevante wie Dysphagie, Kiefer Verschiebung und Gewichtsverlust.

Krebserkrankungen in der Kopf- und Nackenregion werden immer häufiger. Dennoch gibt es ein begrenztes Verständnis der Tumormikroumgebung und der Mechanismen der Behandlungsresistenz in dieser Region. Diese Technik kann verwendet werden, um die native Mikroumgebung von Kopf-Hals-Tumoren auf zugängliche Weise zu rekapitulieren und produziert klinische Manifestationen ähnlich denen beim Menschen.

Wenn die Tumorzellkultur 70% Konfluenz erreicht hat, waschen Sie die Zellen dreimal mit kalten PBS pro Waschgang und befestigen Sie die Zellen mit genügend 0,25% Trypsin, um die untere Oberfläche des Kulturkolbens zu bedecken. Nach drei bis vier Minuten im Zellkultur-Inkubator die Ablösung unter einem Lichtmikroskop bestätigen und das Trypsin mit 12 MilliliterDM F12 Medium neutralisieren, ergänzt durch fetales Rinderserum. Übertragen Sie die Zellsuspension in ein 50 Milliliter konisches Rohr und mischen Sie die Zellen drei- bis viermal durch Inversion.

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