Mouse NIH 3T3 transformed with Moloney murine leukaemia mutants
Description de la lignée cellulaire
From a set of packaging cell lines useful for generating helper-free recombinant retroviruses with ectotropic host ranges. Two mutant Moloney murine leukaemia virus-derived proviral genomes carrying complementary mutations in the gag-pol or env regions were sequentially introduced into NIH 3T3 cells by co-transformation. Each genome contained a deletion of the Psi sequence necessary for the efficient encapsidation of retroviral genomes into virus particles and additional alterations at the 3′end of the provirus. Psi-CRE can be used to isolate clones that stably produce high titres (1000,000 cfu/ml) of recombinant retroviruses with ectotropic host ranges, viral producers derived from Psi-CRE do not transfer the packaging function or yield helper virus.
Application
Ecotropic helper-free recombinant retroviral packaging cell line. Valuable for `in vivo′ gene transfer studies aimed at cell lineage analysis and development of human gene replacement therapy.
Milieu de culture
DMEM + 2mM Glutamine + 10% Foetal Bovine Serum (FBS) (NB. The results obtained by the depositor were gained using 10% Calf Serum (CS)).
Procédure de repiquage
Split sub-confluent cultures (70-80%) 1:3 to 1:6 i.e. seeding at 2-4 x 10,000 cells/cm2 using 0.25% trypsin or trypsin/EDTA; 5-10% CO2, 37°C.
Autres remarques
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