If one is performing some experiments with cancerous cell lines, will it be a correct strategy to use HEK 293T as a normal cell line as a control?
As a point of clarification, HEK293 cells are immortalized by Ad 5 E1A and E1B. HEK293T cells also express SV40 large T antigen. Thus, the cells lack normal pRb and p53 functions. I would never consider these cells to be 'normal'.
Since Hek293T originate from Kidney (but with unknown cellular origin), you might choose something more suiteable for each type of your cancerous cell lines. They (the Heks) might be even of neuronal origin. They do not have the normal karyotype, they are transformed with the help of Adenos and (as consequence) large T ag positive. So, they are not really normal.
Hi Debasish, you can use HEK cells as "normal"/healthy/non-malignant control cells depending on your experimental approaches.
However, apoptotic mechanisms in HEK cells are totally dysregulated (that is reason why HEK cells often applied in transfection experiments), so you should use other cells for analysis of apoptotic effects or assessement of apoptotic read-out.
As a point of clarification, HEK293 cells are immortalized by Ad 5 E1A and E1B. HEK293T cells also express SV40 large T antigen. Thus, the cells lack normal pRb and p53 functions. I would never consider these cells to be 'normal'.
In alternative to HEK cells, you can use a fibroblast cell line as a control.
Basically , 293T CELLS should be use as a test tube and I would be very cautious to explore any biological function with these cells.
As Didier states 293's are just test tubes you can grow on a dish. Nothing about them approaches 'normal'.
see
the 293 Variant Viewer, http://www.hek293genome.org/index.php
and the corresponding nature communications paper
HEK293 cell line was frequently used as non- or low tumorigenic for testing oncogenic properties of cancer-associated genes. However, wild type 293 cells or the empty vector transfected 293 cells demonstrated the ability to form colonies in soft agar and tumors of different size with varying frequencies in immunocompromized mice. 293 cells have highly aberrant karyotype and constitutively express Ad5 E1A/E1B proteins, which deregulate pRB/p53pathways. 293T cell line in addition expresses a temperature-sensitive allele of the SV40 T antigen and thereby is even more tumorigenic. Thus, HEK 293T cell line cannot be used as a normal cells (from “HEK293 cell line in cell biology and cancer research: origin, phenotype, karyotype, tumorigenicity”, manuscript in preparation).
Even if cells are not isolated from a cancerous tissue, the act of immortalisation for continued culture would mean every culture line is not normal and depending on your definition of cancer could also be considered cancer-like. Very few cells continually proliferate passage after passage, stem cells being the exception and even those have their limits. However whether those cells would form tumor or gone on to metastasise is place in an in vivo setting is another thing. So again it depends what you define as cancerous, the unchecked cell proliferation or the ability to cause damage in vivo.
You may want to consider ordering primary cells from Lonza/ATCC. You effectively only get one or no passages out of them but it would be considered more normal. Or if you have animal facility a cheaper option would to just culture the primary cells yourself from a spare animal but I am guessing this takes some time to set up and optimise.
Otherwise I would choose a cell line that has a low growth-rate, maybe NHDF cell. The biology controlling its cell-cycle is likely to be 'more normal'.
Hi,
The 293T cell line is a highly transfectable derivative of human embryonic kidney 293 cells and contains the SV40 T-antigen.
HEK293 is embryonic kidney immortalized cell line, which represents the epithelial morphology and possibility to form tumors in nude mice.
HEK 293 cells are popular for their ease of growth and transfection, making them a common cell culture in cancer research. HEK 293 can be employed as a highly malignant tumor model as its tumorigenicity increases significantly in high-passage. The tumorigenicity of the HEK 293 cell line reached 100% when the passage exceeded 65.
1. Shen C, Gu M, Song C, Miao L, Hu L, Liang D, Zheng C., The tumorigenicity diversification in human embryonic kidney 293 cell line cultured in vitro., Biologicals. 2008 Jul;36(4):263-8. doi: 10.1016/j.biologicals.2008.02.002. Epub 2008 Apr 18.
2. Stepanenko AA, Dmitrenko VV2., HEK293 in cell biology and cancer research: phenotype, karyotype, tumorigenicity, and stress-induced genome-phenotype evolution., Gene. 2015 Sep 15;569(2):182-90. doi: 10.1016/j.gene.2015.05.065. Epub 2015 May 27.
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