HEK 293 cells

Human embryonic kidney 293 cells, also often referred to as HEK 293, HEK-293, 293 cells, or less precisely as HEK cells, are a specific cell line originally derived from human embryonic kidney cells grown in tissue culture taken from a female fetus. HEK 293 cells have been widely used in cell biology research for many years, because of their reliable growth and propensity for transfection. They are also used by the biotechnology industry to produce therapeutic proteins and viruses for gene therapy.


293FT cells, a variation of 293 cell

HEK 293 cells were generated in 1973 by transfection of cultures of normal human embryonic kidney cells with sheared adenovirus 5 DNA in Alex van der Eb's laboratory in Leiden, the Netherlands. The cells were obtained from a single, aborted or miscarried fetus, the precise origin of which is unclear.[1][2] The cells were cultured by van der Eb; the transduction by adenovirus was performed by Frank Graham, a post-doc in van der Eb's lab. They were published in 1977 after Graham left Leiden for McMaster University.[3] They are called HEK since they originated in human embryonic kidney cultures, while the number 293 came from Graham's habit of numbering his experiments; the original HEK 293 cell clone was from his 293rd experiment. Graham performed the transfection a total of eight times, obtaining just one clone of cells that were cultured for several months. After presumably adapting to tissue culture, cells from this clone developed into the relatively stable HEK 293 line.

Subsequent analysis has shown that the transformation was brought about by inserting ~4.5 kilobases from the left arm of the viral genome, which became incorporated into human chromosome 19.[4]

For many years it was assumed that HEK 293 cells were generated by transformation of either a fibroblastic, endothelial or epithelial cell, all of which are abundant in kidneys. However, the original adenovirus transformation was inefficient, suggesting that the cell that finally produced the HEK 293 line may have been unusual in some fashion. Graham and coworkers provided evidence that HEK 293 cells and other human cell lines generated by adenovirus transformation of human embryonic kidney cells have many properties of immature neurons, suggesting that the adenovirus preferentially transformed a neuronal lineage cell in the original kidney culture.[5]

A comprehensive study of the genomes and transcriptomes of HEK 293 and five derivative cell lines compared the HEK 293 transcriptome with that of human kidney, adrenal, pituitary and central nervous tissue.[6] The HEK 293 pattern most closely resembled that of adrenal cells, which have many neuronal properties. Given the location of the adrenal gland (adrenal means "next to the kidney"), a few adrenal cells could plausibly have appeared in an embryonic kidney derived culture, and could be preferentially transformed by adenovirus. Adenoviruses transform neuronal lineage cells much more efficiently than typical human kidney epithelial cells.[5] An embryonic adrenal precursor cell therefore seems the most likely origin cell of the HEK 293 line. As a consequence, HEK 293 cells should not be used as an in vitro model of typical kidney cells.

HEK 293 cells have a complex karyotype, exhibiting two or more copies of each chromosome and with a modal chromosome number of 64. They are described as hypotriploid, containing less than three times the number of chromosomes of a haploid human gamete. Chromosomal abnormalities include a total of three copies of the X chromosomes and four copies of chromosome 17 and chromosome 22.[6][7] The presence of multiple X chromosomes and the lack of any trace of Y chromosome derived sequence suggest that the source fetus was female.


Multiple variants of HEK 293 have been reported.[citation needed]

  • HEK 293F
  • HEK 293FT
  • HEK 293T
  • HEK 293S
  • HEK 293FTM
  • HEK 293SG
  • HEK 293SGGD
  • HEK 293H
  • HEK 293E
  • HEK EBNA1-6E[8]
  • HEK 293MSR
  • HEK 293A


Immunofluorescent HEK 293 cells

HEK 293 cells are straightforward to grow in culture and to transfect. They have been used as hosts for gene expression. Typically, these experiments involve transfecting in a gene (or combination of genes) of interest, and then analyzing the expressed protein. The widespread use of this cell line is due to its transfectability by the various techniques, including calcium phosphate method, achieving efficiencies approaching 100%.

Examples of such experiments include:

HEK 293 cells were adapted to grow in suspension culture, as opposed to proliferation on plastic plates, in 1985.[14] This enabled the growth of large amounts of recombinant adenovirus vectors.

A more specific use of HEK 293 cells is in the propagation of adenoviral vectors.[15] Viruses offer an efficient means of delivering genes into cells, which they evolved to do, and are thus of great use as experimental tools. However, as pathogens, they also present a risk to the experimenter. This danger can be avoided by the use of viruses which lack key genes, and which are thus unable to replicate after entering a cell. In order to propagate such viral vectors, a cell line that expresses the missing genes is required. Since HEK 293 cells express a number of adenoviral genes, they can be used to propagate adenoviral vectors in which these genes (typically, E1 and E3) are deleted, such as AdEasy.[16] However, homologous recombination between the inserted cellular Ad5 sequence and the vector sequence, although rare, can restore the replication capacity to the vector.[17]

An important variant of this cell line is the 293T cell line. It contains the SV40 large T-antigen that allows for episomal replication of transfected plasmids containing the SV40 origin of replication. This allows for amplification of transfected plasmids and extended temporal expression of desired gene products. HEK 293, and especially HEK 293T, cells are commonly used for the production of various retroviral vectors.[18] Various retroviral packaging cell lines are also based on these cells.

Native proteins of interestEdit

Depending on various conditions, the gene expression of HEK 293 cells may vary. The following proteins of interest (among many others) are commonly found in untreated HEK 293 cells:


Alvin Wong argues that despite the uncertainty over the origin of the fetus used to obtain the cell line, circumstantial evidence strongly suggests that it came from an elective abortion. In the context of Catholic teaching, this presents ethical difficulties for using HEK 293 and derivative products, such as vaccines.[23][24]

The US Conference of Catholic Bishops, with the document receiving approval from the Pope, have stated that “it is morally acceptable to receive COVID-19 vaccines that have used cell lines from aborted fetuses in their research and production process”, while at the same time calling on governments and vaccine producers to stop using these cell lines.[25]

Pro-life medical ethicists have said that since the cell lines are immortal, receiving the COVID-19 vaccine does not increase demand for abortion.[citation needed]

During the COVID-19 pandemic, anti-vaccination activists noted that HEK 293 cells are used in the manufacturing of the Oxford–AstraZeneca COVID-19 vaccine (AKA AZD1222). The cells are filtered out of the final products.[26][24]


  1. ^ van der Eb A. "USA FDA CTR For Biologics Evaluation and Research Vaccines and Related Biological Products Advisory Committee Meeting" (PDF). Lines 14–22: USFDA. p. 81. Archived from the original (PDF) on 2017-05-16. Retrieved August 11, 2012.CS1 maint: location (link)
  2. ^ Austriaco N (May 25, 2020). "Moral Guidance on Using COVID-19 Vaccines Developed with Human Fetal Cell Lines". Public Discourse. Witherspoon Institute. Retrieved December 23, 2020. I received an e-mail a few months ago from Professor Frank Graham, who established this cell line. He tells me that to the best of his knowledge, the exact origin of the HEK293 fetal cells is unclear. They could have come from either a spontaneous miscarriage or an elective abortion. Regardless, the abortions that gave rise to the three cell lines—or in the possible case of HEK293, the miscarriage—happened decades ago.
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  4. ^ Louis N, Evelegh C, Graham FL (July 1997). "Cloning and sequencing of the cellular-viral junctions from the human adenovirus type 5 transformed 293 cell line". Virology. 233 (2): 423–9. doi:10.1006/viro.1997.8597. PMID 9217065.
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  6. ^ a b Lin YC, Boone M, Meuris L, Lemmens I, Van Roy N, Soete A, et al. (September 2014). "Genome dynamics of the human embryonic kidney 293 lineage in response to cell biology manipulations". Nature Communications. 5 (8): 4767. Bibcode:2014NatCo...5.4767L. doi:10.1038/ncomms5767. PMC 4166678. PMID 25182477.
  7. ^ "ECACC Catalogue Entry for HEK 293". hpacultures.org.uk. ECACC. Archived from the original on 2012-05-02. Retrieved 2012-03-18.
  8. ^ "HEK293 expression platform (L-10894/11266/11565)" (PDF). National Research Council Canada.
  9. ^ Fredj S, Sampson KJ, Liu H, Kass RS (May 2006). "Molecular basis of ranolazine block of LQT-3 mutant sodium channels: evidence for site of action". British Journal of Pharmacology. 148 (1): 16–24. doi:10.1038/sj.bjp.0706709. PMC 1617037. PMID 16520744.
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  11. ^ Kanno T, Yamamoto H, Yaguchi T, Hi R, Mukasa T, Fujikawa H, et al. (June 2006). "The linoleic acid derivative DCP-LA selectively activates PKC-epsilon, possibly binding to the phosphatidylserine binding site". Journal of Lipid Research. 47 (6): 1146–56. doi:10.1194/jlr.M500329-JLR200. PMID 16520488.
  12. ^ Li T, Paudel HK (March 2006). "Glycogen synthase kinase 3beta phosphorylates Alzheimer's disease-specific Ser396 of microtubule-associated protein tau by a sequential mechanism". Biochemistry. 45 (10): 3125–33. doi:10.1021/bi051634r. PMID 16519507.
  13. ^ Mustafa H, Strasser B, Rauth S, Irving RA, Wark KL (April 2006). "Identification of a functional nuclear export signal in the green fluorescent protein asFP499". Biochemical and Biophysical Research Communications. 342 (4): 1178–82. doi:10.1016/j.bbrc.2006.02.077. PMID 16516151.
  14. ^ Stillman BW, Gluzman Y (August 1985). "Replication and supercoiling of simian virus 40 DNA in cell extracts from human cells". Molecular and Cellular Biology. 5 (8): 2051–60. doi:10.1128/mcb.5.8.2051. PMC 366923. PMID 3018548.
  15. ^ Thomas P, Smart TG (2005). "HEK293 cell line: a vehicle for the expression of recombinant proteins". Journal of Pharmacological and Toxicological Methods. 51 (3): 187–200. doi:10.1016/j.vascn.2004.08.014. PMID 15862464.
  16. ^ He TC, Zhou S, da Costa LT, Yu J, Kinzler KW, Vogelstein B (March 1998). "A simplified system for generating recombinant adenoviruses". Proceedings of the National Academy of Sciences of the United States of America. 95 (5): 2509–14. Bibcode:1998PNAS...95.2509H. doi:10.1073/pnas.95.5.2509. PMC 19394. PMID 9482916.
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  18. ^ Fanelli A (2016). "HEK293 Cell Line: human embryonic kidney cells". Retrieved 3 December 2017.
  19. ^ Dautzenberg FM, Higelin J, Teichert U (February 2000). "Functional characterization of corticotropin-releasing factor type 1 receptor endogenously expressed in human embryonic kidney 293 cells". European Journal of Pharmacology. 390 (1–2): 51–9. doi:10.1016/S0014-2999(99)00915-2. PMID 10708706.
  20. ^ Meyer zu Heringdorf D, Lass H, Kuchar I, Lipinski M, Alemany R, Rümenapp U, Jakobs KH (March 2001). "Stimulation of intracellular sphingosine-1-phosphate production by G-protein-coupled sphingosine-1-phosphate receptors". European Journal of Pharmacology. 414 (2–3): 145–54. doi:10.1016/S0014-2999(01)00789-0. PMID 11239914.
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  25. ^ "Covid: Vatican says coronavirus vaccines 'morally acceptable'". BBC. 22 December 2020.
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