Abstract
The mouse macrophage-like cell line RAW264.7, the most commonly used mouse macrophage cell line in medical research, was originally reported to be free of replication-competent murine leukemia virus (MuLV) despite its origin in a tumor induced by Abelson MuLV containing Moloney MuLV as helper virus. As currently available, however, we find that it produces significant levels of ecotropic MuLV with the biologic features of the Moloney isolate and also MuLV of the polytropic or MCF class. Newborn mice developed lymphoma following inoculation with the MuLV mixture expressed by these cells. These findings should be considered in interpretation of increasingly widespread use of these cells for propagation of other viruses, studies of biological responses to virus infection and use in RNA interference and cell signalling studies.
Highlights
In contrast to most other mouse-derived cell cultures, the macrophage-like cell line RAW264.7 [1] supports replication of murine noroviruses and is widely used for this purpose [2]
The RAW264.7 cell line was derived about 30 years ago from a tumor developing in a BAB/14 mouse, a BALB/c IgH congenic strain, inoculated with Abelson murine leukemia virus (MuLV), a defective transforming virus containing the vabl tyrosine kinase oncogene, and replication-competent Moloney (Mo-MuLV) that served as helper virus [1]
To look for expression of MuLV-encoded protein, RAW264.7 cell pellets were fixed in formalin and embedded in paraffin; after antigen retrieval using proteinase K, sections were stained by avidin-biotin immunohistochemistry (IHC) using groupreactive goat anti-Rauscher MuLV p30 antibody
Summary
In contrast to most other mouse-derived cell cultures, the macrophage-like cell line RAW264.7 [1] supports replication of murine noroviruses and is widely used for this purpose [2]. In tests for the presence of infectious MuLV, cell-free harvests of RAW264.7 supernatants from two separate acquisitions from ATCC were positive using the XC plaque assay [7] in SC-1 cells [8], indicating the presence of virus of the ecotropic MuLV class. Negative results with mAbs reactive with xenotropic MuLVs indicated no significant population of this class in RAW264.7 supernatants (data not shown).
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