HEK cell
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Human Embryonic Kidney cells, also known as HEK cells, HEK 293 or just 293 cells, are an epithelial cell line originally derived, as their name indicates, from embryonic human kidney. HEK cells are not themselves particularly interesting, but are very easy to work with, and so are a widely-used cell line in cell biology research. It should be noted that cultures of the original (untransformed) HEK line have become contaminated with HeLa cells, which have displaced the cell line as originally established.
[edit] Origins of HEK Cells
HEK 293 cells were generated by transformation of cultures of normal human embryonic kidney cells with sheared adenovirus 5 DNA in the laboratory of Frank Graham in the late 1970s (Graham 1977).
Subsequent analysis has shown that the transformation was brought about by an insert consisting of ~4.5 kilobases from the left arm of the viral genome (Louis 1997).
[edit] Uses of HEK 293 Cells
As an experimentally transformed cell line, HEK cells are not a particularly good model for normal cells, cancer cells, or any other kind of cell that is a fundamental object of research. However, they are extremely easy to work with, being straightforward to culture and to transfect, and so can be used in experiments in which the behaviour of the cell itself is not of interest. Typically, these experiments involve transfecting in a gene (or combination of genes) of interest, and then analysing the expressed protein; essentially, the cell is used simply as a test tube with a membrane. The widespread use of this cell line is due to its extreme transfectability by the calcium phosphate method, achieving efficiencies approaching 100% as determined by FACS using a 2XBBS buffer. A lower efficiency might be achievable with an HBS buffer.
An important variant of this cell line is the 293T cell line that contains, in addition, 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 the desired gene products. Note that any similarly domesticated cell line can be used for this sort of work; HeLa, COS and Chinese Hamster Ovary cell are common alternatives.
Examples of such experiments include:
- A study of the effects of a drug on sodium channels [1]
- Testing of an inducible RNA interference system [2]
- Testing of an isoform-selective protein kinase C agonist [3]
- Investigation of the interaction between two proteins [4]
- Analysis of a nuclear export signal in a protein [5]
A more specific use of HEK cells is in the propagation of adenoviral vectors. Viruses offer an extremely efficient means of delivering genes into cells, since this is what they have evolved to do, and are thus of great use as experimental tools. However, as pathogens, they also present a degree of danger 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 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 (He 1998).
293, and especially 293T, cells are commonly used for the production of lentiviral and retroviral vectors. Various retroviral and lentiviral packaging cell lines are based on these cells.
[edit] External links
- Maintaining HEK Cell Cultures
- Graham et al., J. Gen Virol 1977 Jul;36(1):59-74
- Cloning and sequencing of the cellular-viral junctions from the human adenovirus type 5 transformed 293 cell line - N. Louis, C. Evelegh, F. L. Graham, Virology 1997 Jul 7;233(2):423-9
- A simplified system for generating recombinant adenoviruses - T. C. He et al., Proc Natl Acad Sci USA 1998 Mar 3;95(5):2509-14
- HEK 293 Database
- 293 Cells in the ATCC database
- The AdEasy website (WARNING - blink tags)
- Transcript of FDA meeting
