4 replies to this topic

[Kirstenf]

Dear Forum

I have recently received 4 cell lines from a collaborator. Unfortunately, 3 of them were contaminated with mycoplasma.

It is 3 very important cell lines for me and throwing them away (which I would prefer) is not an option. I would like to hear the forums opinion on rescuing cell lines by treating them with gentamycin. What would the recommended procedure be (dose, time)?

I know this a bit of a “Jack in a box” to open this discussion. As far as I can see, it has not been discussed in the forum previously. And I’m looking forward to getting you input.

Kirsten
Department of Haematology
Aalborg Hospital, Denmark

[warsel]

Kirstenf, on Jul 17 2009, 01:23 AM, said:

Dear Forum

I have recently received 4 cell lines from a collaborator. Unfortunately, 3 of them were contaminated with mycoplasma.

It is 3 very important cell lines for me and throwing them away (which I would prefer) is not an option. I would like to hear the forums opinion on rescuing cell lines by treating them with gentamycin. What would the recommended procedure be (dose, time)?

I know this a bit of a “Jack in a box” to open this discussion. As far as I can see, it has not been discussed in the forum previously. And I’m looking forward to getting you input.

Kirsten
Department of Haematology
Aalborg Hospital, Denmark

I did gentamycin treatment before and that worked for me (meaning cells divided normally again and did not look like they had the infection any more by DAPI stain), although I have to say I did not feel good about using that cell line - maybe irrational though.

[gfischer]

The first thing I would do is request that your collaborator send new, mycoplasma-free, aliquots of cells.  Aggressive antibiotic treatment should work, but you never know what subtle effects it could have on your cells.

[leelee]

It has been covered many a time in this forum actually (if you try searching the 1999-2009 archives you will see dozens of variations of this question).

The short answer is NO treatment is not an option, you must get new uncontaminated cells either from your collaborator or another source (if possible?). You can never be sure you will completely get rid of the infection, and like gfisher said you will never know what effects both the infection and the antibiotics have had on your cells hence you can never be sure of any results you get with said cells.

Your work, your choice- but it seems far too many scientists these days underestimate the significance of this issue!!!

[GeorgeWolff]

Certainly with leelee on this.  
Please contact your colleague - it's possible that they're working in ignorance of the problem.  Don't forget all the cell culture work that was found to be corrupted by HeLa cell contamination in the 60's and 70's.