Cell Culture Services

Department Manager:
Carmel Burns, NB1-25

The primary goal of the Cell Services lab is to provide researchers with healthy viable cells and other specialized cell culture techniques.

The facility is well equipped with a warmroom, coldroom, roller bottle machines and spinner culture vessels. We have the capacity to provide large and small-scale cell cultures. The cells we have in the core are grown in media prepared in the media lab and are routinely tested for mycoplasma and Isoenzyme analysis to ensure they are contaminant free.

Cell culture work will be offered on an individual contractual basis. This will be an agreement between Cell Services personnel and the Researcher. At that time a plan will be drawn up, giving lead-time and estimated cost for the cell service.

As part of our quality control program we have a Mycoplasma and Quarantine facility. The core will test for and eradicate mycoplasma from cell cultures. All cell culture is performed in a sterile environment employing aseptic techniques.

The Cell Culture Core provides Cell Banking to Investigators. The Core has the capacity to freeze and store 10,000 vials.

Any cells we receive for culturing will be quarantined until they are tested and found to be free from mycoplasma.

Mycoplasma Testing

We routinely perform a direct and an indirect method of testing for Mycoplasma. The following 2 tests are done in parallel.

Direct --- The growth of colonies on agar

This method requires the use of enriched mycoplasmal media and controlled conditions. This method does not detect all strains but with the appropriate controls this works very well. Direct culturing is a reliable test and takes 2 weeks to complete.

Indirect --- Boehringer ELISA Kit.

We routinely use an ELISA kit from Boehringer Mannheim. This detects the 4 most common types of mycoplasma to contaminate cells. This is a sensitive and convenient method for testing.

Useful Links

• Roche Molecular Biochemicals: www.IbuyRMB.com

Preparing Your Samples for Mycoplasma Testing

Cells should be grown for at least 3 days without antibiotics. Media should not be changed prior to testing. For monolayers, scrape cells and provide the cell suspension in a 5ml amount. For suspensions, 5mls is sufficient. Results will be sent out 2 weeks later. If the result is positive the researcher will be notified immediately.

Eradication of Mycoplasma from Cell Cultures

The core will clean up cells that are contaminated with mycoplasma. This is a lengthy process, which involves culturing the cells in specific antibiotics. Cells will be grown with and without antibiotics for several weeks and tested regularly. Frozen stocks will be made when the cells are mycoplasma free.

Central Cell Services highly recommends that cells obtained from any outside source such as other laboratories or institutions should be tested for mycoplasma prior to culturing and storage.