Replication of Enterotropic and Polytropic Murine Coronaviruses in Cultured Cell Lines of Mouse Origin.

Abstract

Abstract:To understand the virus-cell interactions that occur during murine coronavirus infection, six murine cell lines (A3-1M, B16, CMT-93, DBT, IC-21 and J774A.1) were inoculated with eight murine coronaviruses, including prototype strains of both polytropic and enterotropic biotypes, and new isolates. All virus strains produced a cytopathic effect (CPE) with cell-to-cell fusion in B16, DBT, IC-21 and J774A.1 cells. The CPE was induced most rapidly in IC-21 cells and was visible microscopically in all cell lines tested. In contrast, the coronaviruses produced little CPE in A3-1M and CMT-93 cells. Although most virus-infected cells, except KQ3E-infected A3-1M, CMT-93 and J774A.1 cells, produced progeny viruses in the supernatants when assayed by plaque formation on DBT cells, the kinetics of viral replication were dependent on both the cell line and virus strain; replication of prototype strains was higher than that of new isolates. There was no significant difference in replication of enterotropic and polytropic strains. B16 cells supported the highest level of viral replication. To determine the sensitivity of the cell lines to murine coronaviruses, the 50% tissue culture infectious dose of the coronaviruses was determined with B16, DBT, IC-21 and J774A.1 cells, and compared to that with DBT cells. The results indicate that IC-21 cells were the most sensitive to murine coronaviruses. These data suggest that B16 and IC-21 cells are suitable for large-scale preparation and isolation of murine coronaviruses, respectively.

Mouse Hepatitis Virus

“…Interest in favoring the neurotropism of some MHV strains for experimental use has promulgated the use of DBT cells and highly specific cell lines, such as OBL21A, an olfactory bulb neuronal cell line derived from transformed neonatal mouse cells (Ryder et al 1990). Recently, B 16 cells, derived from a melanoma cell line of C57BL mice (Silagi 1969), and A3-1M cells, derived from 129/SvJ embryonic stem cells (Kyuwa 1997), have shown utility for isolation and propagation of a variety of MHV strains and isolates (Kyuwa et al 2000). Recently, B 16 cells, derived from a melanoma cell line of C57BL mice (Silagi 1969), and A3-1M cells, derived from 129/SvJ embryonic stem cells (Kyuwa 1997), have shown utility for isolation and propagation of a variety of MHV strains and isolates (Kyuwa et al 2000).…”

“…Prototype MHV strains, which have been passaged many times both in vitro and in vivo, tend to grow well in a variety of cell lines, and this can be true of enterotropic MHV strains that have been likewise manipulated (Kyuwa et al 2000). Passage history of various MHV strains, and the viral genetic variability that is likely to ensue, clearly influences their growth characteristics in vitro, explaining apparent discrepancies in results among different laboratories.…”

“…Passage history of various MHV strains, and the viral genetic variability that is likely to ensue, clearly influences their growth characteristics in vitro, explaining apparent discrepancies in results among different laboratories. The study concluded that the kinetics of virus replication depended upon both the cell line and the virus strain, and that prototype viruses replicated better than new isolates (Kyuwa et al 2000). J774A.1 cells produced high titers of MHV-1, -3, -A59, -JHM, -S, and-DVIM as well, and thus were deemed optimal for growth of a variety of MHV strains .…”

Experimental Evaluation of Cross-contamination between Cryotubes Containing Mouse 2-cell Embryos and Murine Pathogens in Liquid Nitrogen Tanks.

“…Thirty 2-cell embryos derived from in vitro fertilization were cryopreserved in a 1 ml cryotube (#366656, Nunc) by the simple vitrification procedure described previously [8]. MHV strain NuU was propagated and titrated on DBT cells as described previously [6]. One milliliter of the supernatant containing 9.8 × 10 6 plaque-forming units of MHV-NuU was frozen in a cryotube (2732-002, IWAKI) at -80°C.…”