293FT Cell Line
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Citas y referencias (3)
Invitrogen™

293FT Cell Line

La línea celular 293FT es perfecta para generar títulos altos de lentivirus mediante el sistema de expresión lentiviral ViraPower™. LaMás información
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Número de catálogoCantidad
R700071 x 10^7 células
Número de catálogo R70007
Precio (MXN)
-
Cantidad:
1 x 10^7 células
La línea celular 293FT es perfecta para generar títulos altos de lentivirus mediante el sistema de expresión lentiviral ViraPower™. La línea celular 293FT es un aislado clonal altamente transfectable y de rápido crecimiento, derivado de células renales embrionarias humanas y transformado con el antígeno T grande SV40. Cuando un vector de expresión ViraPower™ y la mezcla de envasado para lentiviral ViraPower™ se cotransfectan en células 293FT se producen altos niveles de ARN viral y de proteínas rev y gag⁄pol necesarias para el envasado.

Ventajas
• Rápido crecimiento
• Altamente transfectable
• Produce altos títulos de virus
• Permite niveles muy altos de proteína

Características principales
• La presencia del antígeno T grande SV40 permite que se expresen niveles muy altos de proteínas a partir de vectores que contienen el origen de SV40. De hecho, más altos que en el caso de otras células 293 o de las células COS transformadas por el antígeno T que se utilizan tradicionalmente

El kit incluye
• Línea celular 293FT suministrada en forma de solución madre congelada en 1 ml de 90 % de medio completo y 10 % de dimetilsulfóxido (DMSO)
Para uso exclusivo en investigación. No diseñado para uso diagnóstico o terapéutico.
Especificaciones
Recomendación de mediosSFM (medio sin suero)
N.° de celdas1 x 107
Línea de productosViraPower
Cantidad1 x 10^7 células
Línea de células293FT
EspecieHumano
Unit SizeEach
Contenido y almacenamiento
• Línea celular 293FT (almacenar en nitrógeno líquido)

Preguntas frecuentes

Do 293FT cells lift off the plate during lentivirus production?

If 293FT cells detach shortly after transfection (4 hours to overnight):

- This may be a sign of Lipofectamine 2000 toxicity. Cells may have been plated too sparsely prior to transfection.
- The cells may not have been handled gently enough (these cells have a tendency to lift off easily).
- The cells may have been kept at room temperature for too long.

If cells detach 48 to 72 hours post-transfection:
- If the cells lift off in large sheets, this may be a sign of lentivirus production.

What are syncytia?

Syncytia are large multi-nucleated cells that result from VSV-G-induced fusion with neighboring 293FT producer cells. Syncytia production is indicative of high transfection efficiency and lentivirus production. Keep in mind, though, that the absence of syncytia does not mean that virus will not be produced.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

Why do I have to culture 293FT cells under Geneticin antibiotic selection?

The 293FT cell line stably expresses the SV40 large T antigen from the pCMVSPORT6Tag.neo plasmid that contains the neomycin resistance marker. In order to maintain the plasmid/phenotype, the cells have to be routinely cultured in medium containing Geneticin (G418) antibiotic at a concentration of 500 µg/mL.

Do you recommend a specific FBS for culturing 293FT cells? Which plastic plates do you recommend?

We use Mycoplasma-tested Gibco FBS (Cat. No. 16000-044). We have observed that when 293FT cells are cultured in the presence of this FBS following the instructions in the manual, virus production is better than that obtained with many other serum sources. We use the following plasticware for 293FT cells:

T175-Fisher Cat. No. 10-126-13; this is a Falcon flask with 0.2 µm vented plug seal cap.
T75-Fisher Cat. No. 07-200-68; this is a Costar flask with 0.2 µm vented seal cap.
100 mm plate-Fisher Cat. No. 08-772E; this is a Falcon tissue culture-treated polystyrene plate.
We get excellent adherence on these plates under routine cell culture/maintenance conditions.

What are the morphological changes that I should be looking for in my 293FT cells as signs of lentivirus production?

During lentivirus production, 293FT cells undergo the following morphological changes:
- They become multi-nucleated (syncytia development)
- They start to look like balloons or as if they are about to explode
- They often, but not always, lift off from the surface
- Untransfected 293FT cells leave empty spaces and “pile” up at other spots in the flask

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

View all 293FT Cell Line FAQs

Citations & References (3)

Citations & References
Abstract
The interaction between HIV-1 Gag and human lysyl-tRNA synthetase during viral assembly.
Authors:Javanbakht H, Halwani R, Cen S, Saadatmand J, Musier-Forsyth K, Gottlinger H, Kleiman L,
Journal:J Biol Chem
PubMed ID:12756246
'Human lysyl-tRNA synthetase (LysRS) is a tRNA-binding protein that is selectively packaged into HIV-1 along with its cognate tRNALys isoacceptors. Evidence exists that Gag alone is sufficient for the incorporation of LysRS into virions. Herein, using both in vitro and in vivo methods, we begin to map regions in Gag ... More
RIP140-targeted repression of gene expression in adipocytes.
Authors:Christian M, Kiskinis E, Debevec D, Leonardsson G, White R, Parker MG,
Journal:Mol Cell Biol
PubMed ID:16227589
'Ligand-dependent repression of nuclear receptor activity forms a novel mechanism for regulating gene expression. To investigate the intrinsic role of the corepressor RIP140, we have monitored gene expression profiles in cells that express or lack the RIP140 gene and that can be induced to undergo adipogenesis in vitro. In contrast ... More
A novel method for long term bone marrow culture and genetic modification of murine neutrophils via retroviral transduction.
Authors:Zemans RL, Briones N, Young SK, Malcolm KC, Refaeli Y, Downey GP, Worthen GS,
Journal:J Immunol Methods
PubMed ID:19010330
'Neutrophils are a critical component of the innate immune response to invading microbial pathogens. However, an excessive and/or prolonged neutrophil response can result in tissue injury that is thought to underlie the pathogenesis of various inflammatory diseases. The development of novel therapeutic strategies for inflammatory diseases depends on an improved ... More