Mammalian cells, e.g., CHO, BHK, HEK293, HT-1080, and NS0 cells, represent important manufacturing platforms in bioengineering. They are widely used for the production of recombinant therapeutic proteins, vaccines, anticancer agents, and other clinically relevant drugs. HEK293 (human embryonic kidney 293) cells and their derived cell lines provide an attractive heterologous system for the development of recombinant proteins or adenovirus productions, not least due to their human-like posttranslational modification of protein molecules to provide the desired biological activity. Secondly, they also exhibit high transfection efficiency yielding high-quality recombinant proteins. They are easy to maintain and express with high fidelity membrane proteins, such as ion channels and transporters, and thus are attractive for structural biology and electrophysiology studies. In this article, we review the literature on HEK293 cells regarding their origins but also stress their advancements into the different cell lines engineered and discuss some significant aspects which make them versatile systems for biopharmaceutical manufacturing, drug screening, structural biology research, and electrophysiology applications.

1.
Amarouch, M.Y., N. Syam, H. Abriel (2013) Biochemical, single-channel, whole-cell patch clamp, and pharmacological analyses of endogenous TRPM4 channels in HEK293 cells. Neurosci Lett 541: 105-110.
2.
Bloom, F.R., P. Price, G. Lao, J.L. Xia, J.H. Crowe, J.R. Battista, R.F. Helm, S. Slaughter, M. Potts (2001) Engineering mammalian cells for solid-state sensor applications. Biosens Bioelectron 16: 603-608.
3.
Büssow, K. (2015) Stable mammalian producer cell lines for structural biology. Curr Opin Struct Biol 32: 81-90.
4.
Casademunt, E., K. Martinelle, M. Jernberg, S. Winge, M. Tiemeyer, L. Biesert, S. Knaub, O. Walter, C. Schröder (2012) The first recombinant human coagulation factor VIII of human origin: human cell line and manufacturing characteristics. Eur J Haematol 89: 165-176.
5.
Croset, A., L. Delafosse, J.P. Gaudry, C. Arod, L. Glez, C. Losberger, D. Begue, A. Krstanovic, F. Robert, F. Vilbois, L. Chevalet, B. Antonsson B (2012) Differences in the glycosylation of recombinant proteins expressed in HEK and CHO cells. J Biotechnol 161: 336-348.
6.
Dalton, A.C., W.A. Barton (2014) Over-expression of secreted proteins from mammalian cell lines. Protein Sci 23: 517-525.
7.
do Amaral, R.L.F., A. de Sousa Bomfim, M.S. de Abreu-Neto, V. Picanço-Castro, E.M. de Sousa Russo, D.T. Covas, K. Swiech (2016) Approaches for recombinant human factor IX production in serum-free suspension cultures. Biotechnol Lett 38: 385-394.
8.
Dumont, J., D. Euwart, B. Mei, et al. (2016) Human cell lines for biopharmaceutical manufacturing: history, status, and future perspectives. Crit Rev Biotechnol 36: 1110-1122.
9.
Eglen, R., T. Reisine (2011) Primary cells and stem cells in drug discovery: emerging tools for high-throughput screening. Assay Drug Dev Technol 9: 108-124.
10.
El Hader, C., S. Tremblay, N. Solban, et al. (2005) HCaRG increases renal cell migration by a TGF-α autocrine loop mechanism. Am J Physiol Renal Physiol 289: F1273-F1280.
11.
Fernández, F.J., M.C. Vega (2013) Technologies to keep an eye on: alternative hosts for protein production in structural biology. Curr Opin Struct Biol 23: 365-373.
12.
Fliedl, L., J. Grillari, R. Grillari-Voglauer (2015) Human cell lines for the production of recombinant proteins: on the horizon. N Biotechnol 32: 673-679.
13.
Geisse, S., C. Fux (2009) Recombinant protein production by transient gene transfer into mammalian cells. Methods Enzymol 463: 223-238.
14.
Grieger, J.C., S.M. Soltys, R.J. Samulski (2016) Production of recombinant adeno-associated virus vectors using suspension HEK293 cells and continuous harvest of vector from the culture media for GMP FIX and FLT1 clinical vector. Mol Ther 24: 287-297.
15.
Gstraunthaler, G (2003) Alternatives to the use of fetal bovine serum: serum-free cell culture. Altex 20: 275-281.
16.
Hacker, D.L., S. Balasubramanian (2016) Recombinant protein production from stable mammalian cell lines and pools. Curr Opin Struct Biol 38: 129-136.
17.
He, Y., K. Wang, N. Yan (2014) The recombinant expression systems for structure determination of eukaryotic membrane proteins. Protein Cell 5: 658-672.
18.
Hutnick, N.A., D.G. Carnathan, S.A. Dubey, et al. (2010) Vaccination with Ad5 vectors expands Ad5-specific CD8+ T cells without altering memory phenotype or functionality. PLoS One 5: e14385.
19.
Jäger, V., K. Büssow, A. Wagner, et al. (2013) High level transient production of recombinant antibodies and antibody fusion proteins in HEK293 cells. BMC Biotechnol 13: 52.
20.
Jaluria, P., K. Konstantopoulos, M. Betenbaugh, et al. (2008) Egr1 and Gas6 facilitate the adaptation of HEK-293 cells to serum-free media by conferring enhanced viability and higher growth rates. Biotechnol Bioeng 99: 1443-1452.
21.
Jayapal, K.P., K.F. Wlaschin, W. Hu, et al. (2007) Recombinant protein therapeutics from CHO cells - 20 years and counting. CHO Consortium, SBE Special Section, https://pdfs.semanticscholar.org/5f96/12ce9170571f296b75246e80cb671bbb886c.pdf.
22.
Kim, K.S., M.S. Kim, J.H. Moon, et al. (2009) Enhancement of recombinant antibody production in HEK 293E cells by WPRE. Biotechnol Bioproc E 14: 633-638.
23.
Kobayashi, N. (2006) Cell therapy for diabetes mellitus. Cell Transplant 15: 849-854.
24.
Kontoravdi, C., N.J. Samsatli, N. Shah (2013) Development and design of bio-pharmaceutical processes. Curr Opin Chem Eng 2: 435-441.
25.
Kuang, H.B., C.L. Miao, W.X. Guo, et al. (2009) Dickkopf-1 enhances migration of HEK293 cell by beta-catenin/E-cadherin degradation. Front Biosci 14: 20.
26.
Kwon, Y.J., C.A. Peng (2001) Impact of cell growth morphology on retroviral transduction: effect of contact inhibition. Biotechnol Prog 17: 240-246.
27.
Lin, Y.C., M. Boone, L. Meuris, et al. (2014) Genome dynamics of the human embryonic kidney 293 lineage in response to cell biology manipulations. Nat Commun 5: 4767.
28.
Louis, N., C. Evelegh, F.L. Graham (1997) Cloning and sequencing of the cellular-viral junctions from the human adenovirus type 5 transformed 293 cell line. Virology 233: 423-429.
29.
Mesnage, R., B. Bernay, G.E. Séralini (2013) Ethoxylated adjuvants of glyphosate-based herbicides are active principles of human cell toxicity. Toxicology 313: 122-128.
30.
Murakami, M., H. Ugai, M. Wang, et al. (2010) An adenoviral vector expressing human adenovirus 5 and 3 fiber proteins for targeting heterogeneous cell populations. Virology 407: 196-205.
31.
Nadeau, I., A. Kamen (2003) Production of adenovirus vector for gene therapy. Biotechnol Adv 20: 475-489.
32.
Nettleship, J.E., R. Assenberg, J.M. Diprose, et al. (2010) Recent advances in the production of proteins in insect and mammalian cells for structural biology. J Struct Biol 172: 55-65.
33.
Ooi, A., A. Wong, L. Esau, et al. (2016) A guide to transient expression of membrane proteins in HEK-293 cells for functional characterization. Front Physiol 7: 300.
34.
Peiperl, L., C. Morgan, Z. Moodie, et al. (2010) Safety and immunogenicity of a replication-defective adenovirus type 5 HIV vaccine in Ad5-seronegative persons: a randomized clinical trial (HVTN 054). PLoS One 5: e13579.
35.
Peng, Z. (2005) Current status of Gendicine in China: recombinant human Ad-p53 agent for treatment of cancers. Hum Gene Ther 16: 1016-1027.
36.
Pham, P.L., S. Perret, H.C. Doan, et al. (2003) Large-scale transient transfection of serum-free suspension-growing HEK293 EBNA1 cells: peptone additives improve cell growth and transfection efficiency. Biotechnol Bioeng 84: 332-342.
37.
Picanco-Castro, V., R. Tage Biaggio, D. Tadeu Cova, et al. (2013) Production of recombinant therapeutic proteins in human cells: current achievements and future perspectives. Protein Pept Lett 20: 1373-1381.
38.
Piyathilaka, M.A., M.M. Pathmalal, K.H. Tennekoon, B.G. De Silva, S.R. Samarakoon, S. Chanthirika (2015) Microcystin-LR-induced cytotoxicity and apoptosis in human embryonic kidney and human kidney adenocarcinoma cell lines. Microbiology 161(Pt 4): 819-828.
39.
Portolano, N., P.J. Watson, L. Fairall, et al. (2014) Recombinant protein expression for structural biology in HEK 293F suspension cells: a novel and accessible approach. J Vis Exp 92: e51897.
40.
Pradhan, A.K., N. Pradhan, P. Mohapatra, et al. (2014) Cytotoxic effect of microbial biosurfactants against human embryonic kidney cancerous cell: HEK-293 and their possible role in apoptosis. Appl Biochem Biotechnol 174: 1850-1858.
41.
Quennville, Y.D., Y. Durocher (2006) Optimized large-scale transient gene expression with a serum-free suspension-growing HEK293 cell line. Canada, BR Institute, p 1.
42.
Robert, M.A., P.S. Chahal, A. Audy, et al. (2017) Manufacturing of recombinant adeno-associated viruses using mammalian expression platforms. Biotechnol J 12.
43.
Román, R., J. Miret, F. Scalia, A. Casablancas, M. Lecina, J.J. Cairó (2016) Enhancing heterologous protein expression and secretion in HEK293 cells by means of combination of CMV promoter and IFNα2 signal peptide. J Biotechnol 239: 57-60.
44.
Rosario, M., A. Bridgeman, E.D. Quakkelaar, et al. (2010) Long peptides induce polyfunctional T cells against conserved regions of HIV-1 with superior breadth to single-gene vaccines in macaques. Eur J Immunol 40: 1973-1984.
45.
Rose, P.W., A. Prlić, A. Altunkaya, et al. (2017) The RCSB Protein Data Bank: integrative view of protein, gene and 3D structural information. Nucleic Acids Res 45: D271-D281.
46.
Rose, P.W., A. Prlić, C. Bi, et al. (2015) The RCSB Protein Data Bank: views of structural biology for basic and applied research and education. Nucleic Acids Res 43: D345-D356.
47.
Scott, C.W., M.F. Peters, Y.P. Dragan (2013) Human induced pluripotent stem cells and their use in drug discovery for toxicity testing. Toxicol Lett 219: 49-58.
48.
Senatore, A., A.N. Boone, J.D. Spafford (2011) Optimized transfection strategy for expression and electrophysiological recording of recombinant voltage-gated ion channels in HEK-293T cells. J Vis Exp 47: 2314.
49.
Shen, C.F., D. Jacob, T. Zhu, et al. (2016) Optimization and scale-up of cell culture and purification processes for production of an adenovirus-vectored tuberculosis vaccine candidate. Vaccine 34: 3381-3387.
50.
Shinde, V., P. Sureshkumar, I. Sotiriadou, et al. (2016) Human embryonic and induced pluripotent stem cell based toxicity testing models: future applications in new drug discovery. Curr Med Chem 23: 3495-3509.
51.
Shukla, S.J., R. Huang, C.P. Austin, M. Xia (2010) The future of toxicity testing: a focus on in vitro methods using a quantitative high-throughput screening platform. Drug Discov Today 15: 997-1007.
52.
Sroka, J., A. Antosik, J. Czyż, et al. (2007) Overexpression of thioredoxin reductase 1 inhibits migration of HEK-293 cells. Biol Cell 99: 677-687.
53.
Stepanenko, A.A., V.V. Dmitrenko (2015) HEK293 in cell biology and cancer research: phenotype, karyotype, tumorigenicity, and stress-induced genome-phenotype evolution. Gene 569: 182-190.
54.
Strzyz, P. (2017) Synthetic biology: designer cells tackle diabetes. Nat Rev Mol Cell Biol 18: 69.
55.
Subedi, G.P., R.W. Johnson, H.A. Moniz, et al. (2015) High yield expression of recombinant human proteins with the transient transfection of HEK293 cells in suspension. J Vis Exp 106: e53568.
56.
Swiech, K., V. Picanço-Castro, D.T. Covas (2012) Human cells: new platform for recombinant therapeutic protein production. Protein Expr Purif 84: 147-153.
57.
Swiech, K., V. Picanço-Castro, D.T. Covas (2017) Production of recombinant coagulation factors: are humans the best host cells? Bioengineered 8: 462-470.
58.
Thomas, P., T.G. Smart (2005) HEK293 cell line: a vehicle for the expression of recombinant proteins. J Pharmacol Toxicol Methods 51: 187-200.
59.
Van der Valk, J., D. Brunner, K. De Smet, et al. (2010) Optimization of chemically defined cell culture media - replacing fetal bovine serum in mammalian in vitro methods. Toxicol In Vitro 24: 1053-1063.
60.
Varki, A. (1993) Biological roles of oligosaccharides: all of the theories are correct. Glycobiology 3: 97-130.
61.
Venereo-Sanchez, A., R. Gilbert, M. Simoneau, et al. (2016) Hemagglutinin and neuraminidase containing virus-like particles produced in HEK-293 suspension culture: an effective influenza vaccine candidate. Vaccine 34: 3371-3380.
62.
Walsh, G., R. Jefferies (2006) Post-translational modifications in the context of therapeutic proteins. Nat Biotechnol 24: 1241-1252.
63.
Wold, W.S.M., K. Toth (2013) Adenovirus vectors for gene therapy, vaccination and cancer gene therapy. Curr Gene Ther 13: 421-433.
64.
Wu, J., K.D. Chen, M. Gao, et al. (2015) Genetic stability of a recombinant adenovirus vaccine vector seed library expressing human papillomavirus type 16 E6 and E7 proteins. Exp Ther Med 9: 1161-1165.
65.
Xie, M., H. Ye, H. Wang, et al. (2016) β-Cell-mimetic designer cells provide closed-loop glycemic control. Science 354: 1296-1301.
66.
Ye, H., M. Xie, S. Xue, et al. (2016) Self-adjusting synthetic gene circuit for correcting insulin resistance. Nat Biomed Eng 1: 0005.
67.
Yu, H., M. Li, W. Wang, et al. (2016) High throughput screening technologies for ion channels. Acta Pharmacol Sin 37: 34-43.
68.
Zhang, B., X. Peng, G. Li, et al. (2015) Oxidative stress is involved in patulin induced apoptosis in HEK293 cells. Toxicon 94: 1-7.
69.
Zhang, Q., H. Xu, S. Zheng, M. Su, J. Wang (2015) Genotoxicity of mesoporous silica nanoparticles in human embryonic kidney 293 cells. Drug Test Anal 7: 787-796.
70.
Zhao, J., X. Qi, Q. Dai, et al. (2017) Toxicity study of ochratoxin A using HEK293 and HepG2 cell lines based on microRNA profiling. Hum Exp Toxicol 36: 8-22.
71.
Zhao, L., L. Fan, X. Zhang, et al. (2007) The role of microenvironment in aggregation of the 293-human embryonic kidney cells. Kor J Chem Eng 24: 796-799.
72.
Zhao, Y., B. Bishop, J.E. Clay, et al. (2011) Automation of large scale transient protein expression in mammalian cells. J Struct Biol 175: 209-215.
73.
Zhu, J. (2012) Mammalian cell protein expression for biopharmaceutical production. Biotechnol Adv 30: 1158-1170.
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