The Cell culture facility is equipped with a warm room, cold room, spinner vessels and roller bottle apparatus. The core can provide specialized cell cultures such as insect cells, suspension and monolayer cultures, and EBV transformations. The cell core has a mycoplasma and quarantine facility. The core can test for and eradicate mycoplasma from cell cultures.
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.
Since each cell request may be different it is best to discuss this in advance with cell services personnel and an estimated cost and lead-time will be given.
The Cell Culture Lab can provide immortalization of Lymphocytes with EBV (Epstein Barr Virus). This is a reliable method to immortalize mammalian cells. The Cell Culture Lab has successfully developed this technique over the last 10 years. Immortalization of cells takes 4-6 weeks to complete. A sample of 12-15 mls of whole blood collected in a heparin tube is required to get started. After 4-6 weeks the customer will be provided with a T25 flask as a growing culture and 4 frozen vials.
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.
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.
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.
The Cell Culture Core has an 8'x10' warmroom. We have 2 Roller-Bottle machines, which can accommodate 80 large roller bottles. The cell culture core provides large-scale spinner cultures, both suspension and monolayer.
Lerner Research Institute
Cleveland Clinic, Mail Code NB21
9500 Euclid Avenue
Cleveland, Ohio 44195
