History of HEK293 Cells
“As we look ahead to a possible COVID-19 vaccine, it’s important to also take a moment to recognize the invaluable contributions that Dr. Graham has made to such an undertaking with his development of the HEK 293 cell line. The NRC is proud to formally recognize and celebrate Dr. Graham’s outstanding contributions to science.”
—Mr. Iain Stewart, President of the National Research Council of Canada
September 25, 2020 – Ottawa, Ontario – National Research Council of Canada
“I take great satisfaction from the fact that the HEK 293 cell line that I created in 1973 has contributed significantly to advances in the fields of gene therapy and development of vaccines. HEK 293 cells have become one of the most commonly used mammalian cell lines both in academic research and in the biotechnology and pharmaceutical industries.”
– Dr. Frank Graham, Professor Emeritus and Distinguished University Professor, McMaster University
In 1972, Dr. Graham, working as a postdoctoral fellow in the lab of Dutch scientist Dr. Alex van der Eb, developed a new method, called calcium phosphate transfection, for introducing DNA into eukaryotic cells. (1-5) Using this technique, he was able to determine the size and location of the transforming region of Human Adenovirus 5 (Ad5), now known as Early Region I, (6) and in 1973 went on to generate the cell line called HEK 293, which contains and expresses the Ad5 transforming genes. (7-10)
On his return to Canada, Dr. Graham continued to characterize the HEK 293 cell line and, in collaboration with his students and colleagues at McMaster University, he further modified the cell line (11-13) in parallel with the development of numerous Ad5-based viral vectors for gene transfer and potential recombinant viral vaccines. Both HEK 293 cells and reagents for construction of Ad vectors were widely distributed by Dr. Graham to the scientific community for studies on gene therapy and vaccine development.
The HEK 293 cell line has since undergone various modifications in laboratories across the globe and has been employed in countless applications. It has been used in the development of numerous Adenovirus vectors, (14-17) some of which have generated effective Ad vector based vaccines, (13, 18-23) e.g. against Ebola, against rabies and more recently against SARS CoV2 (COVID-19). The cells are also used extensively in the testing and the production of a variety of medicines and in gene therapy, especially in production of AAV based gene transfer vectors.
In 2020, The National Research Council of Canada (NRC) formally recognized the scientific impact of the HEK 293 cell line, originally created by Dr. Frank Graham nearly 50 years ago, and its importance in the production of biologic molecules, including current COVID-19 vaccine product candidates. https://www.canada.ca/en/national-research-council/news/2020/09/recognizing-the-scientific-impact-of-dr-frank-grahams-hek-293-cell-line.html
References
1. Graham, F.L., van der Eb., A.J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 52, 456-467, 1973.
2. Graham, F.L., van der Eb, A.J. Transformation of rat cells by DNA of human adenovirus 5. Virology 54, 536-539, 1973
3. Graham, F.L., Veldhuisen, G., and Wilkie, N.M. Infectious Herpesvirus DNA. Nature New Biology 245, 265-266, 1973
4. Bacchetti, S., and Graham, F.L. Transfer of the gene for thymidine kinase to thymidine kinase-deficient human cells by purified herpes simplex viral DNA. Proc. Natl. Acad. Sci. U.S.A. 74, 1509-1594, 1977
5. van der Eb, A.J., and Graham, F.L. Assay of transforming activity of tumor virus DNA. Methods in Enzymology: Nucleic Acids and Protein Synthesis. Vol. 65 (eds. L. Grossman and K. Moldave). Academic Press, N.Y., 1980.
6. Graham, F.L., van der Eb, A.J., and Heijneker, J.L. Size and location of the transforming region in human adenovirus type 5 DNA. Nature 251, 687-691, 1974
7. Graham, F.L., Smiley, J., Russell, W.C., and Nairn, R. Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J. Gen. Virol. 36, 59-72, 1977
8. Graham. F.L Growth of 293 cells in suspension culture. J. Gen. Virol. 68: 937-940, 1987
9. Louis, N., Evelegh, C. and Graham, F. L. Cloning and sequencing of the cellular/viral junctions from the human adenovirus type 5 transformed 293 cell line. Virology, 233:423-429, 1997
10. Shaw, G., Morse, S., Ararat, M. and Graham, F.L. Preferential transformation of human cells by human adenoviruses and the origins of HEK 293 cells. FASEB J., 16: 869-871, 2002
11. Krougliak, V. and Graham, F. L. Development of cell lines capable of complementing E1, E4 and protein IX defective adenovirus type 5 mutants. Human Gene Therapy 6: 1575-1586, 1995
12. Chen, L., Anton, M. and Graham, F. L. Production and characterization of human 293 cell lines expressing the site-specific recombinase Cre. Somat. Cell and Molec. Genet. 22: 477-488, 1996
13. Matthews, D. A., Cummings, D., Evelegh, C., Graham, F. L. and Prevec, L. Development of a modified 293 cell line expressing the lac repressor and its use in the rescue of recombinant adenoviruses expressing high levels of rabies glycoprotein (G). J. Gen. Virol. 80: 345-53, 1999
14. Graham, F.L. and Prevec, L. Manipulation of adenovirus vectors. Methods in Molecular Biology, Vol. 7: Gene Transfer and Expression Techniques. Eds. E.J. Murray and J.M. Walker. Humana Press Inc., Clifton, N.J. pp 109-128, 1991
15. Bett, A. J., Prevec, L., and Graham, F. L Packaging capacity and stability of human adenovirus type 5 vectors. J. Virol. 67: 5911- 5921, 1993
16. Bett, A. J., Haddara, W., Prevec, L. and Graham, F.L An efficient and flexible system for construction of adenovirus vectors with insertions or deletions in early regions 1 and 3. Proc. Natl. Acad. Sci. US 91: 8802-8806, 1994
17. Hitt, M., Ng, P. and Graham, F. L Construction and propagation of human adenovirus vectors. Cell Biology: A Laboratory Handbook Ed. J. E. Celis. Academic Press. 3rd Edition, pp 435-443, 2005
18. McDermott, M.R., Graham, F.L., Hanke, T. and Johnson, D.C Protection of mice against lethal challenge with herpes simplex virus by vaccination with an adenovirus vector expressing HSV glycoprotein B. Virology 169: 244-247, 1989
19. Prevec, L., Schneider, M., Rosenthal, K.L., Belbeck, L.W., Derbyshire, J.B. and Graham, F.L. Use of human adenovirus based vectors for antigen expression in animals. J. Gen. Virol. 70: 429-434, 1989
20. Graham, F.L Adenoviruses as expression vectors and recombinant vaccines. Trends in Biotechnology 8: 85-87, 1990
21. Prevec, L., Campbell, J.B., Christie, B.S., Belbeck, L. and Graham, F.L A recombinant human adenovirus vaccine against rabies. J. Inf. Dis. 161: 27-30, 1990
22. See, R.H., Petric, M., Zakhartchouk, A.N., Lawrence, D.J., Mok, C. P.Y., Hogan, R. J., Rowe, T., Zitzow,L.A., Karunakaran ,K. P., Hitt, M.M., Graham, F. L., Prevec,L., Mahony, J. B., Sharon, C. Auperin, T., Rini, J.N. Tingle, A.J., Scheifele, D.W., Skowronski, D.M., Patrick, D.M., Babiuk, L.A., Gauldie, J., Voss, T.G., Roper, R.L., Brunham, R.C. and Finlay, B.B.. Comparative evaluation of two severe acute respiratory syndrome (SARS) vaccine candidates in mice challenged with SARS-Coronavirus. J. Gen. Virol. 87: 641-650, 2006
23. Wang, J., Thorson, L., Stokes, R.W., Santosuosso, M., Huygen, K., Zganiacz, A., Hitt, M., and Xing, Z. Single mucosal, but not parenteral, immunization with recombinant adenoviral-based vaccine provides potent protection from pulmonary tuberculosis. J. Immunol. 173: 6357-65, 2004