M. dunni (Clone III8C)

Detection of replication-competent retroviruses in gene therapy.

Derived from the tail tissue of a normal female mouse, Mus dunni. The cells were cloned at passage 14 using Tererasiki plates. M. dunni (Clone III8C) supports the replication of all four classes of murine leukaemia viruses (MuLV); ecotropic, amphotropic, xenotropic and mink cell focus-forming viruses (MCF), showing cytopathic effect. The cells lack most endogenous murine leukaemia virus-related sequences. They are resistant to infection by Moloney MuLV, due to post-translational modifications of the ecotropic MuLV receptor. Confluent cultures will remain more firmly attached in RPMI 1640 or DMEM + 10% FBS during virus assays.

Mouse tail, normal fibroblast

McCoy′s 5a + 2mM Glutamine + 10% Foetal Bovine Serum (FBS).

Split sub-confluent cultures (70-80%) 1:3 to 1:8 i.e. seeding at 1-4x10,000 cells/cm² using 0.25% trypsin or trypsin/EDTA; 5% CO₂; 37°C.

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