ABSTRACT
Guinea pigs (Cavia porcellus) have been used in research since the 19th century to collect data due to their physiological similarities to humans. Today, animals perform a vital role in experiments and concerns for laboratory animal welfare are enshrined in the 3R framework of reduction, refinement and replacement. This case study explores a refinement in humane euthanasia of guinea pigs via the use of an irreversible penetrating spring-loaded captive bolt (CB). Penetrating spring-loaded CB stunning for euthanasia (CBE) was performed on 12 guinea pigs with the parameters for humane slaughter of production animals in order to assess the suitability of this method of euthanasia in contrast to blunt force trauma (BFT). All 12 of the guinea pigs were rendered immediately unconscious with excellent experimental tissue quality collection, high repeatability of results and operator (n = 8) preference over BFT. Overall, CBE in guinea pigs appears to be a feasible refinement for animal welfare, human preference and improved tissue quality for experimental collection in settings where uncontaminated tissues are required.
ABSTRACT
Biopsy is often used to investigate brain tumour-specific abnormalities so that treatments can be appropriately tailored. Dacomitinib (PF-00299804) is a tyrosine kinase inhibitor (TKI), which is predicted to only be effective in cancers where the targets of this drug (EGFR, ERBB2, ERBB4) are abnormally active. Here we describe a method by which serial biopsy can be used to validate response to dacomitinib treatment in vivo using a mouse glioblastoma model. In order to determine the feasibility of conducting serial brain biopsies in mouse models with minimal morbidity, and if successful, investigate whether this can facilitate evaluation of chemotherapeutic response, an orthotopic model of glioblastoma was used. Immunodeficient mice received cortical implants of the human glioblastoma cell line, U87MG, modified to express the constitutively-active EGFR mutant, EGFRvIII, GFP and luciferase. Tumour growth was monitored using bioluminescence imaging. Upon attainment of a moderate tumour size, free-hand biopsy was performed on a subgroup of animals. Animal monitoring using a neurological severity score (NSS) showed that all mice survived the procedure with minimal perioperative morbidity and recovered to similar levels as controls over a period of five days. The technique was used to evaluate dacomitinib-mediated inhibition of EGFRvIII two hours after drug administration. We show that serial tissue samples can be obtained, that the samples retain histological features of the tumour, and are of sufficient quality to determine response to treatment. This approach represents a significant advance in murine brain surgery that may be applicable to other brain tumour models. Importantly, the methodology has the potential to accelerate the preclinical in vivo drug screening process.
