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Procedure for Submitting Cell Cultures for Mycoplasma Testing

1. Passage the cell culture twice in a medium that is free of antibiotics.

2. Prepare sample(s) following one of the procedures below based on cell type.

Adherent Cells
  • Aseptically seed a sterile 25-cm2 flask (T-25) with approximately 5 x 104 cells in 10 mL of medium and grow for 2 population doublings (about 48 hours).
  • Without replacing the current culture medium, aseptically fill the flask completely with fresh culture medium. Seal the lid of the closed flask with Parafilm sealer.
Suspension Cells
  • Count cells.
  • Aseptically place approximately 2 x 105 cells in 10 mL of medium into a sterile container, either a small plastic tube with a screw-cap closure or a T-25 flask. Seal the lid of the closed container with Parafilm sealer.

3. If you are submitting more than one cell culture sample, put samples for each culture in separate zipper-locked or sealed bags. This will protect the integrity of the other sample(s) if one leaks.

4. Fill out the order form that you received after making your appointment; make sure you include your reference number. Pack cell culture sample(s) at room temperature in a sturdy box to prevent breakage. Each flask/tube should be wrapped in plastic bubble wrap or comparable packing material to prevent breakage or leakage. Additional packing material should be used for protection in shipping. Do not use vented caps as they may leak during shipment.

Note: Samples received at ATCC that show signs of leaking or obvious bacteria/fungal contamination will not be tested.

5. Send materials at room temperature by rapid courier to ATCC (maximum two-day delivery). Materials must arrive at ATCC on Monday, Tuesday or by noon on Wednesday of the week of your appointment. Include signed order form with samples(s).

We will send results by fax. DNA stain results will be reported in 7 to 10 work days. Direct culture results will be reported in 4 to 5 weeks.

Note: ATCC does not accept culture sample(s) containing HIV or BSL 3 or 4 agents for mycoplasma testing. ATCC offers Mycoplasma Testing Service for research purposes only. This service has not been validated to support any clinical diagnosis. Testing is not carried out under GLP conditions.

Parafilm is a trademark of Pechiney Plastic Packaging, Inc.