Bioluminescent Orthotopic Model of Pancreatic Cancer Progression

Summary

Improved understanding of pancreatic cancer biology is critically needed to enable the development of better therapeutic options to treat pancreatic cancer. To address this need, we demonstrate an orthotopic model of pancreatic cancer that permits non-invasive monitoring of cancer progression using in vivo bioluminescence imaging.

Abstract

Pancreatic cancer has an extremely poor five-year survival rate of 4-6%. New therapeutic options are critically needed and depend on improved understanding of pancreatic cancer biology. To better understand the interaction of cancer cells with the pancreatic microenvironment, we demonstrate an orthotopic model of pancreatic cancer that permits non-invasive monitoring of cancer progression. Luciferase-tagged pancreatic cancer cells are resuspended in Matrigel and delivered into the pancreatic tail during laparotomy. Matrigel solidifies at body temperature to prevent leakage of cancer cells during injection. Primary tumor growth and metastasis to distant organs are monitored following injection of the luciferase substrate luciferin, using in vivo imaging of bioluminescence emission from the cancer cells. In vivo imaging also may be used to track primary tumor recurrence after resection. This orthotopic model is suited to both syngeneic and xenograft models and may be used in pre-clinical trials to investigate the impact of novel anti-cancer therapeutics on the growth of the primary pancreatic tumor and metastasis.

Introduction

Pancreatic cancer is the fourth leading cause of cancer-related death, with a 5-year survival rate of 4-6%.^1,2 Only 15% of patients are diagnosed early enough in the disease to be eligible for surgery, and tumors recur in >80% of those patients.^3,4 Gemcitabine is used for treatment of pancreatic adenocarcinomas, however, chemoresistance is common and often the drug has little impact on overall survival.⁵ New pharmacological strategies to treat pancreatic cancer are critically needed. Their development depends on significantly improved understanding of the key steps of disease progression that may be sensitive to therapeutic in....

Protocol

The protocol being demonstrated is performed under the guidance and approval of the author's institution's animal care and use committee. All experiments are executed in compliance with all relevant guidelines, regulations and regulatory agencies.

1. Transducing Pancreatic Cancer Cell Lines

1. Transduce pancreatic cancer cells to express luciferase as previously described.^12,13 Panc-1 and Capan-1 pancreatic cancer cell lines transduced with firefly luciferase are used here.

Note: Renilla luciferase or bacterial luciferase may also be used.

2. P....

Results

This method describes an orthotopic model of pancreatic cancer using surgical procedures, including induction of anesthesia, laparotomy, injection of cancer cells in Matrigel and abdominal closure (Figure 1A). The injected cells form a bubble in the surface of the pancreas (Figure 1B). Pancreatic cancer progression may be non-invasively monitored using in vivo bioluminescence imaging to track cancer cell proliferation and dissemination (Figure 2). Liver me.......

Discussion

Here we describe an orthotopic model for longitudinal assessment of pancreatic tumor development and progression. Primary tumor growth kinetics are reproducible (Figure 3) and may be non-invasively monitored using bioluminescence imaging of luciferase-tagged cells, e.g. for analyses of tumor response to novel anti-pancreatic cancer therapeutics. Consistent with the human disease, the model shows local pancreatic invasion (Figure 4A) which allows investigation of tumor cel.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Clean Bench coat
Heating pad			Set to 37 °C
Ivis Lumina ll Bioluminescent imager	Caliper		Alternative bioluminescent imaging systems include In vivo F PRO (Carestream) and Photon Imager (Biospace Lab)
Dissecting scissors
Iris forceps (serrated)
Needle holder
27 G 0.3 ml insulin syringe	Terumo	T35525M2913