This work was supported by the National Health and Medical Research Council, Australia (1008865), the Australian Research Council (LE110100125), the National Cancer Institute (CA138687-01), Erica Sloan is supported by an Early Career Fellowship from the National Breast Cancer Foundation, Australia. Corina Kim-Fuchs is supported by a fellowship from the Swiss Cancer League and an HDR scholarship from Monash Institute of Pharmaceutical Sciences. Eliane Angst is supported by a grant from the Bern Cancer League.

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
<ol>
	<li>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....

<ol>
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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...

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 &gt;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 intervention.
Orthotopic models of pancreatic cancer emulate key aspects of the human disease, making them ideal tools for studying the biology of pancreatic cancer.6-9 In contrast to in vitro cell-based assays of pancreatic cancer cell behavior and subcutaneous in vivo models of pancreatic cancer, orthotopic models allow investigation of tumor cell interactions with the pancreatic microenvironment. The kinetics of disease progression are highly reproducible in orthotopic models and occur over a short time frame (weeks), which makes them well suited to pre-clinical testing of novel therapeutics. This is in contrast to transgenic models where disease onset occurs over a longer and more variable time frame (months to 1 year).10 When used with more aggressive cell lines, orthotopic models of pancreatic cancer have patterns of spontaneous metastasis similar to those seen in patients.8 Expression of bioluminescent reporter genes such as firefly luciferase facilitates longitudinal monitoring of tumor growth, metastatic dissemination, recurrence and response to therapeutics.6,11 
Here we describe an orthotopic model of pancreatic cancer that utilizes Matrigel for localized cell delivery and in vivo bioluminescence imaging for non-invasive monitoring of tumor progression. This orthotopic model of pancreatic cancer allows non-invasive analyses of disease progression and response to therapeutic interventions in syngenic or xenograft models.

<table border="1">
 <tbody>
 <tr>
 <td>Equipment</td>
 <td>Company</td>
 <td>Catalog Number</td>
 <td>Comments</td>
 </tr>
 <tr>
 <td>Clean Bench coat</td>
 <td></td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Heating pad</td>
 <td></td>
 <td></td>
 <td>Set to 37 &deg;C </td>
 </tr>
 <tr>
 <td>Ivis Lumina ll Bioluminescent imager</td>
 <td>Caliper </td>
 <td></td>
 <td>Alternative bioluminescent imaging systems include In vivo F PRO (Carestream) and Photon Imager (Biospace Lab)</td>
 </tr>
 <tr>
 <td>Dissecting scissors</td>
 <td></td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Iris forceps (serrated)</td>
 <td></td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Needle holder</td>
 <td></td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>27 G 0.3 ml insulin syringe</td>
 <td>Terumo</td>
 <td>T35525M2913</td>
 <td></td>
 </tr>
 </tbody>
</table>

bioluminescent orthotopic model of pancreatic cancer progression

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.

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...

Watch this Scientific Journal Video about Bioluminescent Orthotopic Model of Pancreatic Cancer Progression at JoVE.com

Bioluminescent Orthotopic Model of Pancreatic Cancer Progression

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.

Pancreatic cancer has an extremely poor five-year survival rate of 4-6%.  New therapeutic options are critically needed and depend on improved ...

bioluminescent-orthotopic-model-of-pancreatic-cancer-progression

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JoVE Journal

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26.7K visualizações.  Monash University. Uma melhor compreensão da biologia do câncer de pâncreas é criticamente necessário para permitir o desenvolvimento de melhores opções terapêuticas para o tratamento de câncer no pâncreas. Para atender a essa necessidade, nós demonstramos um modelo ortotópico de câncer pancreático que permite o monitoramento não-invasivo de progressão do câncer usando In vivo Imagem de bioluminescência.