Fluorescent Orthotopic Mouse Model of Pancreatic Cancer

Abstract

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 serumfree 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.

Phototoxicity and cell passage affect intracellular reactive oxygen species levels and sensitivity towards non-thermal plasma treatment in fluorescently-labeled cancer cells

“…These considerations should not only be taken into account for experiments utilizing 2D monolayer cultures demonstrated here, but also for those using 3D in vitro models. Moreover, immunofluorescence is also used in in vivo research for tracking tumor growth in orthotopic models and follow-up of metastasis [43,44]. Although ambient light is not a confounding factor in situ in the in vivo environment, our findings could have important implications to take into account during preparatory cell culturing and experimental handling prior to tumor cell inoculation.…”

Ultrasound-Guided Orthotopic Implantation of Murine Pancreatic Ductal Adenocarcinoma

“…, allowing for host mice with additional genetic manipulations that would be time-consuming to breed in to the KPC mouse model. However, pancreatic tumor implantation requires a labor-intensive surgical procedure that introduces aberrant inflammation at the suture site in the abdominal wall24,25,26 , and often includes a lengthy post-operative recovery27,28,29 .…”