Experience with mouse hepatitis virus sanitation in three transplantable murine tumour lines

Transmission of viral infection by tumour lines or other biological materials may have confounding effects on research. Many research organizations require screening for viral agents of all cell lines, tumours, sera and other biologicals before implantation or inoculation into animal models. Screening for viral contamination is done by the mouse antibody production (MAP) test, by cell culture, or alternatively by direct detection of the viral agents by polymerase chain reaction (PCR). The description of procedures for sanitation of infected cell lines or tumours is sparse. The present report describes the procedures used for sanitation of three transplantable murine tumour lines, which were transplanted in vivo in a mouse hepatitis virus (MHV)-infected colony of mice at the Department of Experimental Clinical Oncology (DECO). The tumours were frozen and serially transplanted three times in a quarantine colony of syngenic mice. Serological examination of the mice transplanted with tumours as well as their cage mates in the quarantine colony did not detect any antibodies against MHV. After repeated serial transplantation in seronegative animals, tumour material was frozen and thawed tumours were later used for transplantation into the newly established virus-free colony of mice at DECO. PCR-based detection of MHV did not reveal any contamination of the tumour examined by this technique, indicating that this murine tumour apparently did not transmit MHV or that MHV was eliminated from the tissue so fast after the infection that it could not be transmitted by the tumour tissue. It is concluded that MHV infection of mice with transplantable murine tumours does not necessarily cause the tumours to be contaminated.