Fluorescent Orthotopic Mouse Model of Pancreatic Cancer.
Pancreatic cancer remains one of the cancers for which survival has not improved substantially in the last few decades. Only 7% of diagnosed patients will survive longer than five years. In order to understand and mimic the microenvironment of pancreatic tumors, we utilized a murine orthotopic model of pancreatic cancer that allows non-invasive imaging of tumor progression in real time. Pancreatic cancer cells expressing green fluorescent protein (PANC-1 GFP) were suspended in basement membrane matrix, high concentration, (e.g., Matrigel HC) with serum-free media and then injected into the tail of the pancreas via laparotomy. The cell suspension in the high concentration basement membrane matrix becomes a gel-like substance once it reaches room temperature; therefore, it gels when it comes in contact with the pancreas, creating a seal at the injection site and preventing any cell leakage. Tumor growth and metastasis to other organs are monitored in live animals by using fluorescence. It is critical to use the appropriate filters for excitation and emission of GFP. The steps for the orthotopic implantation are detailed in this article so researchers can easily replicate the procedure in nude mice. The main steps of this protocol are preparation of the cell suspension, surgical implantation, and whole body fluorescent in vivo imaging. This orthotopic model is designed to investigate the efficacy of novel therapeutics on primary and metastatic tumors.
College of Pharmacy, Western University of Health Sciences.
Jonathan A
JA
Moreno
UVP, LLC.
Antonio
A
Sanchez
Department of Surgery, School of Medicine, University of California San Diego; Anticancer Inc.
Robert M
RM
Hoffman
College of Pharmacy, Western University of Health Sciences.
Saima
S
Nur
College of Pharmacy, Western University of Health Sciences; mlambros@westernu.edu.
Maria P
MP
Lambros
D000818:Animals; D016895:Culture Media, Serum-Free; D004195:Disease Models, Animal; D018450:Disease Progression; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D010190:Pancreatic Neoplasms
D016895:Culture Media, Serum-Free; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins
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