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Fungi contamination in cell culture work



Fungi contamination in cell culture work

Fungi contamination is commonly seen in cell culture work. Here is an old discussion thread from bio newsgroup on fungi contamination and use of antibiotics.

We have comme across several cultivations of mammalian cells being "infected" by fibric material. The media we are using is made up from pulver and then sterile filtered, tested for (bacterial) infection. When running continous processes after 2/3 weeks or so we encounter clogging due to some fibres that attached to our reactor inside. Of course we did sterile testing before. To insure we can do the experiments we want to do we add antibiotics, penicillin/streptomycin/neomycin. All material is from GIBCO or Sigma, approved quality.

Is it reasonble to believe that the fungii we saw under the
microscopes/scanning microscopes are imported through the antibiotics ?
As antibiotics are produced from fungii, could it be that during
downstreamprocessing the spores are not totally removed, and end up in the
suspensions ?
Does anyone have had similiar experience/thoughts ?
Should I fight the fungii with antimycotica ?


I always filter-sterilize my media after adding everything,
including the antibiotics if I use them. I never get infected cultures
except when using plates rather than flasks to culture cells. In
plates, an occasional spore gets in when transporting the plates
between the incubator and the hood or scope.
I rarely use antibiotics or anti-mycotics. They should not
be necessary and they often affect the growth of the mammalian cells
as well.
--
********************************************************************
* Brian Foley

If you are using cell culture grade antibiotic solutions from GIBCO,
then it is not reasonable

Last update 01-Jan-2002, Rating of 12 votes.

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By jw on 30-Mar-2008
my cell lines keep getting contaminated with fungi. don't know why? help!
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By sharad on 18-Mar-2008
Is any one using the cell line Kasumi-1. i am facing problem in growing them. if possible contact me.
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By Lalo on 14-Feb-2007
i`m working with fibroblast with iRNA my question is, in my flaks there ir firis material this is a ibdication a fungal contamination, or is a other type of material i the medium .
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By khalid enan on 07-Oct-2006
research assistant in central lab - ministry of science and technology
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By jim on 21-Sep-2006
There is a very good disposable hemacytometer I used for manual cell counting from Cell-VU ® CBC . (www.cellvu.com/drm700directions.htm)
It's cheaper compare with glasses one, and it works well for my cell countings.
It consists of a dual-chamber glass slide, with a printed inert surface. This surface supports two cover slips. The reverse side of cover slip has grid pattern.It consists of 9 large squares.The four corner squares are divided into 16 smaller squares . The central square, used prima-rily for erythrocyte, platelet and sperm counts, is divided into 100 small squares .
I t can be used counting for
A: Erythrocyte count
B: Leukocyte count
C: Platelet count
D: Eosinophil count
E: Spinal/Body Fluids
F: Semen Analysis
G: sperm counts and motility
For detail direction, see Link:
www.cellvu.com/drm700directions.htm


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By peggy on 17-May-2006
I was wondering if any one could tell me how to get a cell count using a hemacytometer?
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By Piyanart on 19-Feb-2006
I recently faced the Hep3B cell culture problems regard to very slow growth rate.

My questions are as follow;
1. I use Dulbecco`s Modified Eagle medium (1X high glucose with L-glutamine, 110 mg/L Sodium pyruvate and pyridoxine hydrochlorite; Invitrogen Cat. No. 11995-065) supplemented with 10% Penicillin-Streptomycin Mixed Solution (Cat. No. 26252-94) and 10% Sodium Pyruvate 100mM (Cat No. 11360-070). This is an appropriate Hep3B cell medium or not?

2. When I detach cell, I use 1 ml of Trysin/EDTA (Cat. No. HK-3120), adding in 10-ml Petri dish for 2-3 minutes. This is also a appropriate condition for Hep3B detachment or not?

3. From time to time, I found fungus and bacteria contamination. How do I solve these problems.

Thank you very for every recommendation

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By Khaled on 17-Jan-2006
I'm using HepG2 cell line in my culture work. I will use DMEM media for culturing. The complete media consists of :
- DMEM
- 10%FBS
- 100 U/ml penicillin/streptomycin.
The concentration of stock solution of penicillin/streptomycin is 10000 U/ml. So, please let me know the volumes of each of these three reagents to prepare 100 ml complete media.
Thanks for help.
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By babu on 05-May-2005
iam student from thisuniversity we were always using antibiotics in the culture media then also we gotinfection tothe culture .what is resion behind this icouldnot got resion for that.but usual practice&well eqiped laboratory conditions can give successful culture.
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By sudeep on 20-Apr-2005
i m working on CHO. i found some black dots in my culture. i found that as bacterial conatamination. i used penicilin and genta. but no effect. the growth of contaminant is slow such that it wont affect cells as well as medium
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By Bhaik on 16-Mar-2005
We are working on BHK21 and L929 cell lines. I am facing problem of severe contamintion, mostly bacterial. I have fumigated the incubator twice in a week, tried with a fresh media also, but still unable to overcome it. Any suggestion please.
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By pooja on 29-Apr-2004
We recently found that unopened bottles of media (from one of the companies that you mentioned) were heavily contaminated with yeast. With that we have reasons to believe that other things from the same company might be contaminated too. It might be a good idea to check your antibiotics before use.
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By samuel on 06-Apr-2004
I am working on HEK 293 CELL lines. I am facing the similar fungal contamination even in flasks and that too after one month of culture establishment.
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By jaspreet on 01-Jan-2002
I am working on mouse neuronal culture. I am facing the similar fungal contamination even in flasks and that too after one month of culture establishment.
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