{"id":20696,"date":"2018-06-25T11:56:00","date_gmt":"2018-06-25T11:56:00","guid":{"rendered":"http:\/\/biomedpharmajournal.org\/?p=20696"},"modified":"2018-10-03T06:51:29","modified_gmt":"2018-10-03T06:51:29","slug":"the-scattered-twelve-tribes-of-hek293","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol11no2\/the-scattered-twelve-tribes-of-hek293\/","title":{"rendered":"The Scattered Twelve Tribes of HEK293"},"content":{"rendered":"

Introduction<\/strong><\/p>\n

Their ease of growth and transfection makes HEK293 cells a common cell culture in academic research. In addition, high transfection efficiency of HEK293 cells enable production of exogenous proteins or viruses for pharmaceutical and biomedical research purposes.1\u20133<\/sup>\u00a0For example, HEK293 cells are frequently used to express potential biological drugs and\/or therapeutic target during the discovery stage.1,4,5\u00a0<\/sup>Recently, HEK293 cells has gained attention due to it is\u00a0 versatility for transfection experiments, particularly the propagation of adenoviral-based and retroviral-based vectors during CART-T bioprocess.6\u20139<\/sup><\/p>\n

A human embryonic kidney cell line was derived from the kidney of a human embryo in Alex van der Eb’s laboratory in\u00a0Netherlands.10<\/sup> After several months of cultivation, the fast-growing variant was established as HEK293 cells. Later, the microarray study demonstrate that HEK293 cells express a multitude of neuron-specific genes, indicating that 293 cells were originated from an immature neuronal cell in the embryonic kidney.10\u00a0<\/sup>From there, many variants of HEK293 cells were developed for specific purposes.11<\/sup>\u00a0Here we review these \u201ctribes\u201d of HEK293 cells that are\u201d scattered\u201d around the world to fulfill their maker\u2019s purpose in cell culture studies.<\/p>\n

The Scattered Twelve Tribes of HEK293<\/strong><\/p>\n

HEK293 – The HEK293 line is the original line established from a primary embryonic human kidney and transformed with sheared human adenovirus type 5 DNA. The E1A adenovirus gene is expressed in these cells and participates in transactivation of some viral promoters, allowing these cells to produce very high levels of protein.2<\/sup><\/p>\n

HEK293S \u2013 HEK293S cell is the original HEK293 line in suspension of modified minimal Eagle\u2019s medium. Full adaptation took about 7 months, and the first passages were so difficult that the few cells that grew through are likely to have been almost clonal. The fully adapted cell line is known as 293S11<\/sup><\/p>\n

HEK293T – \u00a0HEK293T is a HEK293 variant that expresses a temperature-sensitive allele of the SV40 T antigen. This enables the amplification of vectors containing the SV40 ori and thus considerably increases the protein expression levels during transient transfection. SV40 T forms a complex with and inhibits p53, possibly further compromising genome integrity.10,12<\/sup><\/p>\n

HEK293F \u2013 HEK293F is a variant of HEK293 cells. HEK293F cells were cloned from the HEK293 cell line and adapted to commercial medium.<\/p>\n

HEK293FT – HEK293 FT is a fast growing variant of HEK293T. HEK293FT cells were cloned from the HEK293T cell line and adapted to commercial media.13<\/sup>\u00a0HEK293FT is designed for lentiviral production. Similar to HEK293T, the 293FT cells stably express the SV40 large T antigen from the pCMVSPORT6TAg. neo plasmid. Expression of the SV40 large T antigen is controlled by the human cytomegalovirus (CMV) promoter and is high-level and constitutive.13<\/sup><\/p>\n

HEK293FTM – HEK293FTM cell is derived from 293 cells by stable transfection of an FRT-site containing plasmid and of a TetR expression plasmid. The FRT site can be used for fast and easy generation of a stably transfected cell pool by co-transfecting a Flp-InTM expression vector containing a gene of interest and a Flp recombinase expression vector. The 293FTM cells were designed for protein-protein interaction studies.11<\/sup><\/p>\n

HEK293SG – HEK293SG was derived from HEK293S by ethylmethanesulfonate (EMS) induced mutation. A Ricin toxin-resistant clone was then selected to become HEK293SG. The line lacked N-acetylglucosaminyltransferase I activity (encoded by the\u00a0MGAT1<\/em>\u00a0gene) and accordingly predominantly modifies glycoproteins with the Man5<\/sub>GlcNAc2<\/sub>\u00a0N-glycan. HEK293SG is used for the production of homogenously N-glycosylated proteins.11<\/sup><\/p>\n

HEK293SGGD -HEK293SGGlycoDelete cell line (293SGGD) derives from 293SG through transfection with an expression plasmid for a Golgi-targeted form of endoT, an endoglycosidase from the fungus Trichoderma reesei. HEK293SGGD are mainly used for glycosylation study.11<\/sup><\/p>\n

HEK293H \u2013 HEK293H were cloned from HEK293 cells by limiting dilution to select a clone with good adherence during plaque assays.<\/p>\n

HEK293E – HEK293E cells are derived from the HEK293 cell line and used for propagation of plasmids and expression of recombinant proteins in mammalian cells. The cell line expresses the EBNA-1 protein for episomal replication of oriP-harboring plasmids.14,15<\/sup><\/p>\n

HEK293MSR – HEK293MSR cell line is genetically engineered from HEK293 expresses the human macrophage scavenger receptor and strongly adheres to standard tissue culture plates for dependable results.16<\/sup><\/p>\n

HEK293A – HEK293A cell is a subclone of the HEK293 cells with a relatively flat morphology. It facilitates the initial production, amplification and titering of replication-incompetent adenovirus. The cell line contains a stably integrated copy of the E1 gene that supplies the E1 proteins (E1a and E1b) required to generate recombinant adenovirus.17,18<\/sup><\/p>\n

Concluding Remarks<\/strong><\/p>\n

HEK293 and its variants have been the most frequently used cells after HeLa in cell biology studies and after CHO in biotechnology.11<\/sup>\u00a0Despite the widespread and historically long term productive exploitation in cell biology, biotechnology, and cancer research, the designed purposes of these 293 variants were are frequently misused.10,13<\/sup>\u00a0Therefore, we review the HEK293 and its variants in order to produce a comprehensive guild line for the intended purpose of these variants in cell culture studies.<\/p>\n

Reference<\/strong><\/p>\n

    \n
  1. Thomas P & Smart T. G. HEK293 cell line: A vehicle for the expression of recombinant proteins. J. Pharmacol. Toxicol. Methods<\/em> 2005;51:187\u2013200.
    \nCrossRef<\/a><\/li>\n
  2. Stepanenko A. A & Dmitrenko V. V. HEK293 in cell biology and cancer research: Phenotype, karyotype, tumorigenicity, and stress-induced genome-phenotype evolution. Gene<\/em> 2015;569:182\u2013190.
    \n    CrossRef<\/a><\/li>\n
  3. J\u00e4ger V. et al.<\/em> High level transient production of recombinant antibodies and antibody fusion proteins in HEK293 cells. BMC Biotechnol.<\/em> 2013;13.
    \n    CrossRef<\/a><\/li>\n
  4. Watanabe H. et al.<\/em> Heat-evoked activation of TRPV4 channels in a HEK293 cell expression system and in native mouse aorta endothelial cells. J. Biol. Chem.<\/em> 2002;277:47044\u201347051.
    \n    CrossRef<\/a><\/li>\n
  5. Srivastava A, Durocher Y & Gamain B. Expressing full-length functional PfEMP1 proteins in the HEK293 expression system. Methods Mol. Biol.<\/em> 2013;923:307\u2013319.
    \n    CrossRef<\/a><\/li>\n
  6. Zhang C, Liu J, Zhong J. F & Zhang X. Engineering CAR-T cells. Biomark. Res.<\/em>\u00a02017;5:22.
    \n    CrossRef<\/a><\/li>\n
  7. Jackson H. J, Rafiq S & Brentjens R. J. Driving CAR T-cells forward. Nature Reviews Clinical Oncology.<\/em> 2016;13:370\u2013383.
    \n    CrossRef<\/a><\/li>\n
  8. Srivastava S & Riddell S. R. Engineering CAR-T cells: Design concepts. Trends in Immunology<\/em> 2015;36:494\u2013502.
    \n    CrossRef<\/a><\/li>\n
  9. Petiot E, Cuperlovic-Culf M, Shen C. F & Kamen A. Influence of HEK293 metabolism on the production of viral vectors and vaccine. Vaccine.<\/em> 2015;33:5974\u20135981.
    \n    CrossRef<\/a><\/li>\n
  10. Shaw G, Morse S, Ararat M & Graham F. L. Preferential transformation of human neuronal cells by human adenoviruses and the origin of HEK 293 cells. FASEB J.<\/em> 2002;16:869\u201371.
    \n    CrossRef<\/a><\/li>\n
  11. Lin Y. C. et al.<\/em> Genome dynamics of the human embryonic kidney 293 lineage in response to cell biology manipulations. Nat. Commun.<\/em> 2014;5.
    \n    CrossRef<\/a><\/li>\n
  12. Rio D. C. et al.<\/em> A Mammalian Host-Vector System that Regulates Expression and Amplification of Transfected Genes by Temperature Induction. Science<\/em> 2015;227:23\u201328.
    \n    CrossRef<\/a><\/li>\n
  13. Oka Y. et al.<\/em> 293FT cells transduced with four transcription factors (OCT4, SOX2, NANOG, and LIN28) generate aberrant ES-like cells. J. Stem Cells Regen. Med.<\/em> 2010;6:149\u2013156.<\/li>\n
  14. Backliwal G. et al.<\/em> Rational vector design and multi-pathway modulation of HEK 293E cells yield recombinant antibody titers exceeding 1 g\/l by transient transfection under serum-free conditions. Nucleic Acids Res.<\/em> 2008;36.
    \n    CrossRef<\/a><\/li>\n
  15. Rajendra Y, Kiseljak D, Baldi L, Hacker D. L & Wurm F. M. Reduced glutamine concentration improves protein production in growth-arrested CHO-DG44 and HEK-293E cells. Biotechnol. Lett.<\/em> 2012;34:619\u2013626.
    \n    CrossRef<\/a><\/li>\n
  16. Lau T & Schloss P. Differential regulation of serotonin transporter cell surface expression. Wiley Interdisciplinary Reviews: Membrane Transport and Signaling.<\/em> 2012;1:259\u2013268.
    \n    CrossRef<\/a><\/li>\n
  17. Tragoolpua K. et al.<\/em> Generation of functional scFv intrabody to abate the expression of CD147 surface molecule of 293A cells. BMC Biotechnol.<\/em> 2008;8.
    \n    CrossRef<\/a><\/li>\n
  18. Xiong Z. et al.<\/em> Room-temperature, atmospheric plasma needle reduces adenovirus gene expression in HEK 293A host cells. Appl. Phys. Lett.<\/em> 2011;99.
    \n    CrossRef<\/a><\/li>\n<\/ol>\n","protected":false},"excerpt":{"rendered":"

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