title: Welfare Assessment following Heterotopic or Orthotopic Inoculation of Bladder Cancer in C57BL/6 Mice.
abstract: Few studies have assessed whether mice used as cancer models experience pain. Despite this possibility, the usual practice is to withhold analgesics as these are generally viewed as confounding. However, pain also alters cancer progression, so preventing it might not only be beneficial to welfare but also to study validity. Establishing the extent to which different cancer models result in pain is an important first step towards their refinement. We used conditioned place preference (CPP) testing and body-weight and behaviour analyses to evaluate the assumption that heterotopically implanted tumours result in less pain and fewer welfare concerns than those implanted orthotopically. C57Bl/6 mice received MB49Luc luciferase expressing bladder cancer cells or saline implanted subcutaneously or into the bladder. These tumour-bearing or control groups underwent 2 daily 45 minute conditioning trials to saline or morphine (2mg/kg) and then a 15 minute drug-free preference test on day 3 of a 3 day cycle, continuing until the study ended. Tumours were imaged and behaviour data obtained following preference tests. Development of preference for the morphine-paired chamber (morphine-seeking) was determined over time. Heterotopic tumour development had no effect on morphine-seeking, and although the restraint used for heterotopic inoculation caused greater initial weight losses than anaesthesia, these mice steadily gained weight and behaved comparatively normally throughout the study. Orthotopic tumour inoculation caused no initial weight losses, but over the final 7 days these mice became less active and lost more body weight than cancer-free controls. This indicated orthotopic implantation probably caused a more negative impact on welfare or conceivably pain; but only according to the current test methods. Pain could not be confirmed because morphine-seeking in the tumour-bearing groups was similar to that seen in controls. Imaging was not found to be an effective method of monitoring tumour development surpassing manual tumour inspection.
authors.affiliation: Institute of Neuroscience, Comparative Biology Centre, Newcastle University, Newcastle upon Tyne, NE2 4HH, England.
authors.forename: Amy
authors.initials: A
authors.lastname: Miller
authors.affiliation: Research Institute of Pharmaceutical Sciences and Departments of Psychology and Pharmacology, Peabody Building, University of Mississippi, Oxford, MS, 38677, United States of America.
authors.forename: Hannah
authors.initials: H
authors.lastname: Burson
authors.affiliation: MDKN, Wilamowitzweg 11, 37085, Göttingen, Germany.
authors.forename: Ariane
authors.initials: A
authors.lastname: Söling
authors.affiliation: Institute of Neuroscience, Comparative Biology Centre, Newcastle University, Newcastle upon Tyne, NE2 4HH, England.
authors.forename: Johnny
authors.identifier: http://orcid.org/0000-0002-0031-3918
authors.initials: J
authors.lastname: Roughan
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D051379:Mice; D008810:Mice, Inbred C57BL; D009368:Neoplasm Transplantation / Q000379:methods*; D001749:Urinary Bladder Neoplasms / Q000503:physiopathology*