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Cell Biology
ATCC® Number: CRL-1573™    Price: $256.00
Designations: 293 [HEK-293]
Depositors:  FL Graham
Biosafety Level: 2 [CELLS CONTAIN ADENOVIRUS ]
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Homo sapiens (human)
Morphology: epithelial
Source: Organ: embryonic kidney
Cell Type: transformed with adenovirus 5 DNA
Permits/Forms: In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
Restrictions: These cells are distributed for research purposes only. 293 cells, their products, or their derivatives may not be distributed to third parties.
Applications: efficacy testing [92587]
transfection host (Nucleofection technology from Lonza
Roche FuGENE® Transfection Reagents)
viruscide testing [92579]
Receptors: vitronectin, expressed
Tumorigenic: YES
DNA Profile (STR): Amelogenin: X
CSF1PO: 11,12
D13S317: 12,14
D16S539: 9,13
D5S818: 8,9
D7S820: 11,12
THO1: 7,9.3
TPOX: 11
vWA: 16,19
Cytogenetic Analysis: This is a hypotriploid human cell line. The modal chromosome number was 64, occurring in 30% of cells. The rate of cells with higher ploidies was 4.2 %. The der(1)t(1;15) (q42;q13), der(19)t(3;19) (q12;q13), der(12)t(8;12) (q22;p13), and four other marker chromosomes were common to most cells. Five other markers occurred in some cells only. The marker der(1) and M8 (or Xq+) were often paired. There were four copies of N17 and N22. Noticeably in addition to three copies of X chromosomes, there were paired Xq+, and a single Xp+ in most cells.
Age: fetus
Comments: Although an earlier report suggested that the cells contained Adenovirus 5 DNA from both the right and left ends of the viral genome [RF32764], it is now clear that only left end sequences are present. [39768]
The line is excellent for titrating human adenoviruses.
The cells express an unusual cell surface receptor for vitronectin composed of the integrin beta-1 subunit and the vitronectin receptor alpha-v subunit. [23406]
The Ad5 insert was cloned and sequenced, and it was determined that a colinear seqment from nts 1 to 4344 is integrated into chromosome 19 (19q13.2). [39768]
Propagation: ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
The cell line does not adhere to the substrate when left at room temperature for any length of time, therefore, live cultures may be received with the cells detached.The cells will re-attach to the flask over a period of several days in culture at 37C.
Subculturing: Protocol:
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
  3. Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
    Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
  4. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.An inoculum of 2 X 10 (3) to 6 X 10 (3) viable cells/cm2 is recommended.
  6. Incubate cultures at 37°C.6. Subculture when cell concentration is between 6 and 7 X 10(4) cells/cm2.

Subcultivation Ratio: 1:10 to 1:20 weekly.
Medium Renewal: Every 2 to 3 days
Preservation: Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium): ATCC 30-2003
derivative: ATCC CRL-10852
derivative: ATCC CRL-12006
derivative: ATCC CRL-12007
derivative: ATCC CRL-12013
derivative: ATCC CRL-12479
derivative: ATCC CRL-2029
derivative: ATCC CRL-2368
purified DNA:ATCC CRL-1573D
References: 21624: Xie QW, et al. Complementation analysis of mutants of nitric oxide synthase reveals that the active site requires two hemes. Proc. Natl. Acad. Sci. USA 93: 4891-4896, 1996. PubMed: 8643499
21631: Da Costa LT, et al. Converting cancer genes into killer genes. Proc. Natl. Acad. Sci. USA 93: 4192-4196, 1996. PubMed: 8633039
22282: Graham FL, et al. Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J. Gen. Virol. 36: 59-72, 1977. PubMed: 886304
22319: Graham FL, et al. Defective transforming capacity of adenovirus type 5 host-range mutants. Virology 86: 10-21, 1978. PubMed: 664220
22699: Harrison T, et al. Host-range mutants of adenovirus type 5 defective for growth in HeLa cells. Virology 77: 319-329, 1977. PubMed: 841862
23406: Bodary SC, McLean JW. The integrin beta 1 subunit associates with the vitronectin receptor alpha v subunit to form a novel vitronectin receptor in a human embryonic kidney cell line. J. Biol. Chem. 265: 5938-5941, 1990. PubMed: 1690718
27819: Goodrum FD, Ornelles DA. The early region 1B 55-kilodalton oncoprotein of adenovirus relieves growth restrictions imposed on viral replication by the cell cycle. J. Virol. 71: 548-561, 1997. PubMed: 8985383
28301: Loffler S, et al. CD9, a tetraspan transmembrane protein, renders cells susceptible to canine distemper virus. J. Virol. 71: 42-49, 1997. PubMed: 8985321
32283: Hu SX, et al. Development of an adenovirus vector with tetracycline-regulatable human tumor necrosis factor alpha gene expression. Cancer Res. 57: 3339-3343, 1997. PubMed: 9269991
32396: Kolanus W, et al. alphaLbeta2 integrin/LFA-1 binding to ICAM-1 induced by cytohesin-1 a cytoplasmic regulatory molecule. Cell 86: 233-242, 1996. PubMed: 8706128
32490: Stauderman KA, et al. Characterization of human recombinant neuronal nicotinic acetylcholine receptor subunit combinations alpha 2 beta 4, alpha 3 beta 4 and alpha 4 beta 4 stably expressed in HEK293 cells. J. Pharmacol. Exp. Ther. 284: 777-789, 1998. PubMed: 9454827
32514: Bartz SR, et al. Human immunodeficiency virus type 1 cell cycle control: Vpr is cytostatic and mediates G2 accumulation by a mechanism which differs from DNA damage checkpoint control. J. Virol. 70: 2324-2331, 1996. PubMed: 8642659
32726: Sandri-Goldin RM, Hibbard MK. The herpes simplex virus type 1 regulatory protein ICP27 coimmunoprecipitates with anti-sm antiserum, and the C terminus appears to be required for this interaction. J. Virol. 70: 108-118, 1996. PubMed: 8523514
32829: Ansieau S, et al. Tumor necrosis factor receptor-associated factor (TRAF)-1, TRAF-2, and TRAF-3 interact in vivo with the CD30 cytoplasmic domain; TRAF-2 mediates CD30-induced nuclear factor kappa B activation. Proc. Natl. Acad. Sci. USA 93: 14053-14058, 1996. PubMed: 8943059
32893: Zhang J, et al. Dynamin and beta-arrestin reveal distinct mechanisms for G protein-coupled receptor internalization. J. Biol. Chem. 271: 18302-18305, 1996. PubMed: 8702465
32914: Oppermann M, et al. Monoclonal antibodies reveal receptor specificity among G-protein-coupled receptor kinases. Proc. Natl. Acad. Sci. USA 93: 7649-7654, 1996. PubMed: 8755530
32921: Xia Y, et al. Nitric oxide synthase generates superoxide and nitric oxide in arginine-depleted cells leading to peroxynitrite-mediated cellular injury. Proc. Natl. Acad. Sci. USA 93: 6770-6774, 1996. PubMed: 8692893
32925: Zhu X, et al. Cell cycle-dependent modulation of telomerase activity in tumor cells. Proc. Natl. Acad. Sci. USA 93: 6091-6095, 1996. PubMed: 8650224
32971: Uebele VN, et al. Functional differences in Kv1.5 currents expressed in mammalian cell lines are due to the presence of endogenous Kvbeta2.1 subunits. J. Biol. Chem. 271: 2406-2412, 1996. PubMed: 8576199
33003: Abell A, et al. Deletions of portions of the extracellular loops of the lutropin/choriogonadotropin receptor decrease the binding affinity for ovine luteinizing hormone, but not human choriogonadotropin, by preventing the formation of mature cell surface receptor. J. Biol. Chem. 271: 4518-4527, 1996. PubMed: 8626807
33010: Tiberi M, et al. Differential regulation of dopamine D1A receptor responsiveness by various G protein-coupled receptor kinases. J. Biol. Chem. 271: 3771-3778, 1996. PubMed: 8631993
33022: Shahrestanifar M, et al. Studies on inhibition of mu and delta opioid receptor binding by dithiothreitol and N-ethylmaleimide. His223 is critical for mu opioid receptor binding and inactivation by N-ethylmaleimide. J. Biol. Chem. 271: 5505-5512, 1996. PubMed: 8621408
33035: Boring L, et al. Molecular cloning and functional expression of murine JE (monocyte chemoattractant protein 1) and murine macrophage inflammatory protein 1alpha receptors. J. Biol. Chem. 271: 7551-7558, 1996. PubMed: 8631787
33036: Noonberg SB, et al. Evidence of post-transcriptional regulation of U6 small nuclear RNA. J. Biol. Chem. 271: 10477-10481, 1996. PubMed: 8631843
33050: Fox JC, Shanley JR. Antisense inhibition of basic fibroblast growth factor induces apoptosis in vascular smooth muscle cells. J. Biol. Chem. 271: 12578-12584, 1996. PubMed: 8647868
33056: Lee MJ, et al. The inducible G protein-coupled receptor edg-1 signals via the Gi/mitogen-activated protein kinase pathway. J. Biol. Chem. 271: 11272-11279, 1996. PubMed: 8626678
33123: Marchand P, et al. Cysteine mutations in the MAM domain result in monomeric meprin and alter stability and activity of the proteinase. J. Biol. Chem. 271: 24236-24241, 1996. PubMed: 8798668
33137: Arai H, Charo IF. Differential regulation of G-protein-mediated signaling by chemokine receptors. J. Biol. Chem. 271: 21814-21819, 1996. PubMed: 8702980
33138: Huang Q, et al. Substrate recognition by tissue factor-factor VIIa. J. Biol. Chem. 271: 21752-21757, 1996. PubMed: 8702971
33157: Monteclaro FS, Charo IF. The amino-terminal extracellular domain of the MCP-1 receptor, but not the RANTES/MIP-1alpha receptor, confers chemokine selectivity. J. Biol. Chem. 271: 19084-19092, 1996. PubMed: 8702581
33158: Keith DE, et al. Morphine activates opioid receptors without causing their rapid internalization. J. Biol. Chem. 271: 19021-19024, 1996. PubMed: 8702570
39768: Louis N, et al. Cloning and sequencing of the cellular-viral junctions from the human adenovirus type 5 transformed 293 cell line. Virology 233: 423-429, 1997. PubMed: 9217065
61259: Shaw G, et al. Preferential transformation of human neuronal cells by human adenoviruses and the origin of HEK 293 cells. FASEB J. 16: 869-871, 2002. PubMed: 11967234
92579: Standard Test Method for Determining the Virus-Eliminating Effectiveness of Liquid Hygienic Handwash and Handrub Agents Using the Fingerpads of Adult Volunteers. West Conshohocken, PA:ASTM International;ASTM Standard Test Method E 1838-02.
92587: Standard Quantitative Disk Carrier Test Method for Determining the Bactericidal, Virucidal, Fungicidal, Mycobactericidal and Sporicidal Activities of Liquid Chemical Germicides. West Conshohocken, PA:ASTM International;ASTM Standard Test Method E 2197-02.
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