Syngeneic Mouse Orthotopic Allografts to Model Pancreatic Cancer

Constantin Schmitt; Dieter Saur; Stefanie B&#228;rthel; Chiara Falcomat&#224;

doi:10.3791/64253

Syngeneic Mouse Orthotopic Allografts to Model Pancreatic Cancer

JoVE Journal
Cancer Research

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JoVE Journal Cancer Research

Syngeneic Mouse Orthotopic Allografts to Model Pancreatic Cancer

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06:20 min

October 4, 2022

DOI: 10.3791/64253-v

Constantin Schmitt^1,2,3, Dieter Saur^1,2,3,4, Stefanie Bärthel*^1,2,3, Chiara Falcomatà*^1,2,3

¹Division of Translational Cancer Research,German Cancer Research Center (DKFZ) and German Cancer Consortium (DKTK), ²Translational Cancer Research and Institute of Experimental Cancer Therapy, Klinikum rechts der Isar, School of Medicine,Technical University of Munich, ³Center for Translational Cancer Research (TranslaTUM), School of Medicine,Technical University of Munich, ⁴Department of Internal Medicine II, Klinikum rechts der Isar,Technical University of Munich

Overview

This protocol outlines the generation of syngeneic mouse orthotopic allografts of pancreatic ductal adenocarcinoma (PDAC), which mimic the disease's biology and therapeutic responses. These models are essential for preclinical studies due to their rapid and reproducible tumor progression.

Key Study Components

Area of Science

Neuroscience
Oncology
Preclinical Research

Background

PDAC is a challenging cancer with poor prognosis.
Syngeneic models provide a relevant biological context for studying tumor behavior.
These models allow for the assessment of drug responses in an immunocompetent environment.
Reproducibility is crucial for validating experimental results.

Purpose of Study

To establish a reliable method for creating PDAC models.
To facilitate studies on tumor progression and treatment efficacy.
To enhance understanding of PDAC biology in a syngeneic context.

Methods Used

Washing PDAC cells with phosphate buffered saline (PBS).
Detaching cells using trypsin for cell culture.
Resuspending cells in DMEM with fetal bovine serum and antibiotics.
Injecting the cell suspension into the pancreas of syngeneic mice.

Main Results

Successful establishment of PDAC models in vivo.
Demonstrated reproducibility of tumor growth.
Ability to study drug responses in a relevant biological setting.
Insights into the tumor microenvironment and its effects on progression.

Conclusions

Syngeneic mouse models are valuable for PDAC research.
This protocol provides a framework for future studies.
Further research can lead to improved therapeutic strategies.

Frequently Asked Questions

What is PDAC?

Pancreatic ductal adenocarcinoma (PDAC) is a type of cancer that originates in the pancreas and is known for its aggressive nature.

Why use syngeneic models?

Syngeneic models provide a more accurate representation of tumor behavior in an immunocompetent environment, which is crucial for studying cancer therapies.

What are the advantages of this protocol?

The protocol allows for reproducible tumor progression studies and the assessment of drug responses in a biologically relevant context.

Who conducted this research?

The research was conducted by Stefanie Barthel and Chiara Falcomata from the laboratory.

What is the significance of using DMEM with FBS?

DMEM with fetal bovine serum provides essential nutrients and growth factors for the cultured PDAC cells, ensuring their viability and functionality.

How can this research impact cancer treatment?

By understanding tumor progression and drug responses, this research can lead to the development of more effective treatment strategies for PDAC.

Syngeneic mouse orthotopic allografts of pancreatic ductal adenocarcinoma (PDAC) recapitulate the biology, phenotypes, and therapeutic responses of disease subtypes. Owing to their fast, reproducible tumor progression, they are widely used in preclinical studies. Here, we show common practices to generate these models, injecting syngeneic murine PDAC cultures into the pancreas.

This protocol provides reproducible, immunocompetent PDAC in vivo models suitable for genotype, phenotype, and drug response studies. The main advantage of this technique is its ability to help investigate tumor progression with a syngeneic biological tumor microenvironment while maintaining the reproducibility of the experiment. Demonstrating the procedure will be Stefanie Barthel and Chiara Falcomata, both are from our laboratory.

Begin the preparation of the tumor cells by washing the pancreatic ductal adenocarcinoma, or PDAC cells, grown in a T75 culture flask with phosphate buffered saline or PBS. Then add 0.5 milliliters of trypsin for five to 10 minutes to detach from the flask. Re-suspend the detached cells in 10 milliliters of DMEM with 10%fetal bovine serum, or FBS, and 0.1%penicillin streptomycin before transferring the cell suspension to a 15 milliliter tube.

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