In Vitro Technologies

Permits and Restrictions	View Permits
Tissue	embryonic kidney
Cell Type	embryonic
Product Format	frozen
Morphology	rounded
Culture Properties	suspension
Biosafety Level	2 [Contains cells transformed with adenovirus 5 DNA] Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Age	fetus
Storage Conditions	liquid nitrogen vapor phase

Karyotype	The following is based on the parent cell line (ATCC CRL-1573): 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.Note: Cytogenetic instability has been reported in the literature for some cell lines.
Images
Derivation	ATCC CRL-1573 (HEK293) cells were progressively adapted from an anchorage-dependent growth mode to a suspension mode.
Comments	ATCC CRL-1573 (HEK293) cells were progressively adapted from an anchorage-dependent growth mode to a suspension mode. Although an earlier report suggested that the cells contained Adenovirus 5 DNA from both the right and left ends of the viral genome, it is now clear that only left end sequences are present. The Ad5 insert was cloned and sequenced, and it was determined that a colinear segment from nts 1 to 4344 is integrated into chromosome 19 (19q13.2).

Complete Growth Medium	The base medium for this cell line is 293 SFM II (Invitrogen, Catalog No. 11686-029). To make the complete growth medium, add the following component to the base medium: 4mM L-glutamine (final conc.)
Subculturing	Volumes are given for a 75 cm² flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes. Cultures can be maintained by addition of fresh medium. Dilute cultures to a cell concentration between 1 x 10⁵ and 3 x 10⁵ cells/mL. Alternatively, cultures can be established by centrifugation with subsequent resuspension at 1 x 10⁵ and 3 x 10⁵ viable cells/mL. Transfer the suspension to a centrifuge tube. Sharply rap the side of the flask against your hand or a protected surface several times to remove any adherent cells. Resuspend the dislodged cells in 5 mL medium and triturate with a small bore pipette until cell clumps are dispersed. Pool resuspended cells into the centrifuge tube. Centrifuge at 125 xg for 5 to 7 minutes. Remove supernatant and resuspend cell pellet with fresh medium. Do not allow the cell concentration to exceed 1 x 10⁶ cells/mL. Medium renewal: Two to three times weekly
Cryopreservation	Freeze medium: Conditioned growth medium (day 3 to 4 cell conditioned medium collected from HEK-293.2sus cultures during subculture procedure) supplemented with 10% (v/v) DMSO. liquid nitrogen vapor phase
Culture Conditions	Temperature: 37°C Atmosphere: air, 95%; carbon dioxide (CO₂), 5%

STR Profile	Amelogenin: X CSF1PO: 12 D13S317: 12,14 D16S539: 9,13 D5S818: 8 D7S820: 11,12 TH01: 7,9.3 TPOX: 11 vWA: 16,19
Population Doubling Time	approximately 34 hours

Name of Depositor	ATCC
Year of Origin	October 2008
References	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 Harrison T, et al. Host-range mutants of adenovirus type 5 defective for growth in HeLa cells. Virology 77: 319-329, 1977. PubMed: 841862 Graham FL, et al. Defective transforming capacity of adenovirus type 5 host-range mutants. Virology 86: 10-21, 1978. PubMed: 664220 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 Da Costa LT, et al. Converting cancer genes into killer genes. Proc. Natl. Acad. Sci. USA 93: 4192-4196, 1996. PubMed: 8633039 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 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 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 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 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 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 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 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 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 Noonberg SB, et al. Evidence of post-transcriptional regulation of U6 small nuclear RNA. J. Biol. Chem. 271: 10477-10481, 1996. PubMed: 8631843 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 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 Arai H, Charo IF. Differential regulation of G-protein-mediated signaling by chemokine receptors. J. Biol. Chem. 271: 21814-21819, 1996. PubMed: 8702980 Huang Q, et al. Substrate recognition by tissue factor-factor VIIa. J. Biol. Chem. 271: 21752-21757, 1996. PubMed: 8702971 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 Keith DE, et al. Morphine activates opioid receptors without causing their rapid internalization. J. Biol. Chem. 271: 19021-19024, 1996. PubMed: 8702570 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 Loffler S, et al. CD9, a tetraspan transmembrane protein, renders cells susceptible to canine distemper virus. J. Virol. 71: 42-49, 1997. PubMed: 8985321 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 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 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 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.

Name of Depositor

ATCC

Year of Origin

October 2008

References

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

Harrison T, et al. Host-range mutants of adenovirus type 5 defective for growth in HeLa cells. Virology 77: 319-329, 1977. PubMed: 841862

Graham FL, et al. Defective transforming capacity of adenovirus type 5 host-range mutants. Virology 86: 10-21, 1978. PubMed: 664220

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

Da Costa LT, et al. Converting cancer genes into killer genes. Proc. Natl. Acad. Sci. USA 93: 4192-4196, 1996. PubMed: 8633039

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

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

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

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

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

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

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

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

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

Noonberg SB, et al. Evidence of post-transcriptional regulation of U6 small nuclear RNA. J. Biol. Chem. 271: 10477-10481, 1996. PubMed: 8631843

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

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

Arai H, Charo IF. Differential regulation of G-protein-mediated signaling by chemokine receptors. J. Biol. Chem. 271: 21814-21819, 1996. PubMed: 8702980

Huang Q, et al. Substrate recognition by tissue factor-factor VIIa. J. Biol. Chem. 271: 21752-21757, 1996. PubMed: 8702971

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

Keith DE, et al. Morphine activates opioid receptors without causing their rapid internalization. J. Biol. Chem. 271: 19021-19024, 1996. PubMed: 8702570

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

Loffler S, et al. CD9, a tetraspan transmembrane protein, renders cells susceptible to canine distemper virus. J. Virol. 71: 42-49, 1997. PubMed: 8985321

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

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

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

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.

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HEK-293.2sus; Embryonic Kidney; Human (Homo sapiens) Item ID: ATCCRL15733

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