Description: 293T cells serially transfected with synthetic HIV Gag-Pol sequences expressed from a Murine Leukaemia Virus (MLV) vector; HIV tat sequences expressed from a MLV vector; HIV rev sequence expressed from a MLV vector, and amphotropic envelope protein expressed from a plasmid. Finally, Green Fluorescent Protein (GFP) introduced by infection with a HIV expression vector. These cells stably express synthetic HIV Gag-Pol, HIV tat and HIV rev sequences, an amphotropic envelope protein and a GFP expressing HIV vector. The cells therefore produce disabled lentivirus containing GFP gene for efficient transduction of dividing and non-dividing cells. Genetically Modified Organism Class II.

Also Known As:

Species: Human

Tissue: Kidney, embryo

Growth Properties: Adherent

Morphology: Epithelial

Growth Medium: DMEM + 2mM Glutamine + 10% Foetal Bovine Serum (FBS).

Subculturing Procedure: Split sub-confluent cultures (70-80%) 1:4 to 1:10 i.e. seeding at 3-5×10,000 cells/cm² using 0.25% trypsin or trypsin/EDTA; 10% CO2; 37°C. Cells may take up to 7 days to recover after resuscitation. If very few cells survive after 48 hours, it is advisable to wait another 2-7 days. During this time changing the media should be avoided as cells that are less than 10% confluent are extremely sensitive to any form of shock. These cells are more sensitive to pH and temperature shock than normal 293T cells, it is advisable to pre-warm media at 37oC before use. Cells detach easily at room temperature or during transit, therefore growing cultures may be received with cells in suspension. These cells must be frozen in glycerol not DMSO.

Release Conditions: Yes. All customers are required to complete a Material Transfer Agreement

Depositor: Professor Mary Collins, Department of Immunology and Molecular Pathology, University College London

Originator: Yes

References: Ikeda Y et. al., Nat. Biotechnol. (2003) May: 21(5): 569-572

Hyperlink to ECACC Cell Line Data Sheet: 04072115