Are you actually working with a neuronal line?

Friday, September 3, 2010

Many labs are either working with or have HEK293 cells frozen back in their repository. HEK293 embryonic kidney cells were cultured from an aborted fetus by Alex van der Eb, a researcher in Holland. Were you aware that they might not actually be of kidney cells and rather of neuronal origin. The cells also made Frank Graham, a researcher with van der Eb, famous for discovering that you could transfect some DNA if you just toss some calcium phosphate in with them. A scientist by the name of Gerry Shaw was trying to stain 293's with a monoclonal antibody that he created to beta-adrenergic receptor kinase 1 (beta-ARK) which should be expressed in the cells. Wanting to be a proper scientist, Shaw wanted to have a negative control for staining, so he grabbed a neurofilament antibody, which should not stain a kidney cell line right? WTF this and other neurofilament antibodies were able to stain the cells! Scratching your head right, so was Shaw. Using microarray data, it was shown that 293's expression pattern of neuronal genes similar to that of other neuronal cell lines. So if you are wanting an in vitro model for kidney research, you might want to go get another cell line.

Link to Shaw's data

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Thomas Joseph

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Fri, Sep 03, 2010, 6:51 am CDT

Ooops!

Nikkilina
Washington University School of Medicine

Fri, Sep 03, 2010, 6:52 am CDT

Um ... wow. WTH?

Ash

Fri, Sep 03, 2010, 8:17 am CDT

Woah.. just trying to imagine how horrifying this news will be to people using the cell line.

Genomic Repairman

Fri, Sep 03, 2010, 8:33 am CDT

Nikki, its a little disconcerting. Ash, its not really that horrifying, you just have to be cognizant of it. This crap has happened before like the use of Chang liver cells as a model for liver and those cells are not actually liver cells, they're HeLa. A good way to solve cell identification and cell-to-cell contamination issues is STR analysis and the repositories usually have the data on hand for cell lines that they provide too.

Fri, Sep 03, 2010, 9:20 am CDT

I'm surprised it's taken this long for someone to confirm the cell line's origin. That's a little uncomfortable.

Fri, Sep 03, 2010, 9:26 am CDT

Nikki, if you are surprised by that, go back and look up Walter Nelson-Rees and Stanley Gartler were exposing HeLa contamination issues in the 60's and 70's and the field largely tried to turn a blind eye.

Fri, Sep 03, 2010, 9:48 am CDT

I would have hoped we had moved past issues like that. Especially since the tests to analyze this are fairly straightforward and common.

Fri, Sep 03, 2010, 10:09 am CDT

Journals are really driving this, some of them are asking for verification of cell lines so a big thumbs up goes out to those guys.

Gerty-Z

Fri, Sep 03, 2010, 11:14 am CDT

Wikipedia has a list of "contaminated" cell lines with ref's that gives an idea of how pervasive this problem is (that we know of)

Arlenna

Fri, Sep 03, 2010, 12:15 pm CDT

Yeah I had read about this. We use HEK293 cells in my lab, however just as 'vessels' for protein overexpression and chemical studies, and I wanted to know more about them. Pretty crazy, but reminds us that we should really pay attention to what we're working with and why.

Venkat

Guest Comment

Tue, Sep 07, 2010, 2:23 am CDT

MDCK says "Yeah! Come running back to me you suckers!"

Tue, Sep 07, 2010, 8:43 am CDT

Too right Venkat, too right.

Physician Scientst

Guest Comment

Thu, Sep 09, 2010, 6:59 am CDT

This is well-known. 293s are probably JG cells from the kidney. By microarray, they express proteins most like the adrenal cortex.

Thu, Sep 09, 2010, 8:00 am CDT

Phys Scientist you are right this isn't late breaking news but I thought it was interesting since a fair amount of labs work with these cells.

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