HEK 293 cells
Human embryonic kidney 293 cells, also often referred to as HEK 293, HEK-293, 293 cells, are an immortalised cell line derived from HEK cells isolated from a female fetus in the 1970s.[1][2]
The HEK 293 cell line has been widely used in research for decades due to its reliable and fast growth and propensity for
History
HEK 293 cells were generated in 1973 by
Subsequent analysis has shown that the
For many years it was assumed that HEK 293 cells were generated by transformation of either a
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.[7] 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.[6] 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
The 293T cell line was created in Michele Calos's lab at Stanford by stable transfection of the HEK 293 cell line with a plasmid encoding a temperature-sensitive mutant of the SV40 large T antigen; it was originally referred to as 293/tsA1609neo.[9] The first reference to the cell line as "293T" may be its use to create the BOSC23 packaging cell line for producing retroviral particles.[10]
Variants
Multiple variants of HEK 293 have been reported.[11][12]
- HEK 293
- HEK 293F
- HEK 293FT
- HEK 293T
- HEK 293S
- HEK 293FTM
- HEK 293SG
- HEK 293SGGD
- HEK 293H
- HEK 293E
- HEK EBNA1-6E[13]
- HEK 293MSR
- HEK 293A
- HEK293-ENT1KO
HEK 293T
The transfection used to create 293T (involving plasmid pRSV-1609) conferred
The full genome sequences of three different isolates of 293T have been determined. They are quite similar to each other but show detectable divergence from the parental HEK 293 cell line.[15]
HEK293-ENT1KO
This mutant strain does not express of the equilibrative nucleoside transporter ENT1. The gene was knocked out using CRISPR-CAS9 and the cell line retains ENT2 expression.[16]
Applications
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:
- Effects of a drug on sodium channels[17]
- Inducible RNA interference system[18]
- Isoform-selective protein kinase C agonist[19]
- Interaction between two proteins[20]
- Nuclear export signal in a protein[21]
HEK 293 cells were adapted to grow in suspension culture, as opposed to proliferation on plastic plates, in 1985.[22] 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
An important variant of this cell line is the
Native proteins of interest
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:
- Corticotrophin releasing factor type 1 receptor[27]
- EDG5[28]
- Muscarinic acetylcholine receptor M3[29]
Bioethics
Alvin Wong, a catholic bioethicist, argues that despite the uncertainty over the origin of the embryonic cells used to obtain the cell line, one can infer that it came from a voluntary abortion. To some, this may present an ethical dilemma for using HEK 293 and derivative products, such as vaccines and many medications.[31][32][33][34]
On 21 December 2020, the Roman Catholic
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.[36]
Regeneron Pharmaceuticals, the maker of REGN-COV2, a therapeutic antibody cocktail used to alleviate symptoms of patients with COVID-19, did not use HEK 293T cells to produce the antibody cocktail but did use those cells to assess the potency of the drug.[37][32]
In response to ethical concerns of vaccine production, several strategies for clinicians to discuss with their patients have been suggested.[38]
See also
References
- PMID 21605454.
- ^ a b 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.
- ^ 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.
{{cite web}}: CS1 maint: location (link) - PMID 886304.
- PMID 9217065.
- ^ S2CID 6519203.
- ^ PMID 25182477.
- ECACC. Archived from the originalon 2012-05-02. Retrieved 2012-03-18.
- PMID 3031469.
- PMID 7690960.
- PMID 34359846.
- ISSN 0892-6638.
- ^ "HEK293 expression platform (L-10894/11266/11565)" (PDF). National Research Council Canada. April 2019.
- PMID 2981116.
- PMID 25182477.
- ISSN 0892-6638.
- PMID 16520744.
- PMID 16522642.
- PMID 16520488.
- PMID 16519507.
- PMID 16516151.
- PMID 3018548.
- PMID 15862464.
- PMID 9482916.
- PMID 21994701.
- ^ Fanelli A (2016). "HEK293 Cell Line: human embryonic kidney cells". Retrieved 3 December 2017.
- PMID 10708706.
- PMID 11239914.
- PMID 18388243.
- PMID 15972814.
- PMID 17091554.
- ^ a b Schorr I (20 December 2020). "The Facts about the COVID Vaccines and Fetal Cell Lines". National Review.
- ^ "Balkan Insight: Tulpina religioasă a anti-vaccinismului se răspândește în Europa Centrală și de Est - International - HotNews.ro". 20 January 2021.
- ^ RELIGIOUS STRAIN OF ANTI-VAX GROWS IN CEE, balkaninsight.com
- ^ "Nota della Congregazione per la Dottrina della Fede sulla moralità dell'uso di alcuni vaccini anti-Covid-19". press.vatican.va. Retrieved 2024-03-06.
- ^ Rahman G (26 November 2020). "There are no foetal cells in the AstraZeneca Covid-19 vaccine". Full Fact.
- ^ Regalado, Antonio (7 October 2020). "Trump's antibody treatment was tested using cells originally derived from an abortion". MIT Technology Review. Retrieved 2021-09-30.
It's how you want to parse it," says Alexandra Bowie, a Regeneron spokesperson. "But the 293T cell lines available today are not considered fetal tissue
- PMID 34172329.
External links
- HEK 293 Transfection and Selection Data @ Cell-culture Database
- A HEK293 Cell Database Archived 2017-05-11 at the Wayback Machine
- 293 Cells (CRL-1573) Archived 2012-06-22 at the ATCCdatabase
- Transcript of FDA meeting, in which, starting page 77, van der Eb describes in detail the origin of HEK 293 cell
- 293T in the Culture Collections of Public Health England
- Cellosaurus entry for HEK 293 and HEK 293T
- ATCC entry for 293T