Orthotopic Implantation of Murine Pancreatic Cancer Cell Lines

The tumorigenicity of the cell lines was determined after orthotopic implantation of the mouse PC cell lines (UN-KC-6141/UN-KPC-960/UN-KPC-961) into the head of the pancreas after 35 passages. Based on the mice background from where the cell lines were generated, UN-KC-6141 cells (1×10⁶) were injected in C57BL/6 mice, whereas UN-KPC-960 and UN-KPC-961 cells (1×10⁶) were injected in mixed background (i.e. B6.129) mice (N = 7). Mouse PC cells suspended in 50 µl sterile PBS were injected orthotopically using the same procedure described by us [13] (link), [14] (link). Subcutaneous tumor growth was evaluated with the UN-KPC-961 cell line only. 5×10⁶ cells were injected (N = 12) in mixed B6.129 background mice on the lateral chest. Tumor growth was monitored by palpation/Vernier caliper measurements in case of subcutaneous tumors. Throughout the experiment, animals were provided with food and water ad libitum and subjected to a 12-h dark/light cycle. Animal studies were performed in accordance with the U.S. Public Health Service "Guidelines for the Care and Use of Laboratory Animals" under an approved protocol by the University of Nebraska Medical Center Institutional Animal Care and Use Committee (IACUC). After euthanization, pancreatic tumors were dissected out, weighed and fixed in 10% formalin (Fisher Scientific, Fair Lawn, NJ) for H&E staining. Gross metastatic lesions were examined in distant organs and processed for histological analysis.