NCRAD MYCOPLASMA SUBMISSION

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Some Alzheimer's Disease Centers may have cell lines already established for individuals in a family. NCRAD is willing to accept these cell lines, but only if they are free of mycoplasma infection. It is crucial that cell lines maintained at NCRAD are free of mycoplasma infection to prevent cross-contamination. For this reason, it is required that ALL cells are tested for mycoplasma before sending the samples to NCRAD. Because this is such an important issue, the laboratory performing the testing will bill NCRAD. You will not be billed for the testing.

If the family is approved for enrollment, please follow the steps below if your Center will be sending cell lines to NCRAD. Every cell line that will be sent to NCRAD must be tested for mycoplasma. Also, the MTA for shipping to NCRAD must be submitted before sending the cell lines to the repository.

• The laboratory can test either tissue culture supernatant or cells, but supernatant is strongly preferred. The cells must be grown for a minimum of 1 week or 2 passages in antibiotic free medium before submitting the sample.
• Collect 1 to 2 mLs from a couple of different dishes/flasks at least 24 hours after the last medium change at which the cells were greater than 80% confluent.
• Supernatant samples may be submitted frozen or refrigerated. If adherent cells are to be submitted, they MUST be harvested using a cell scraper (no trypsin). All cells should be submitted cryopreserved only after meeting the culturing requirements indicated above. The laboratory greatly prefers supernatant/conditioned medium.
• The samples must be labeled with a unique ID, which may be the family and individual ID or an internal laboratory ID. Before sending the sample(s), complete the sample submission form. If there are any questions or concerns, contact the IU VPF Manager, Troy Hawkins at troyhawk@iupui.edu or (317) 278-1628.
• The address for sending samples will be provided after the sample submission form is filled out and submitted. Someone will contact you to coordinate sending the samples for mycoplasma testing after the submission form is received. Do not send samples without first submitting the form. If samples are received without proper notification they may be rejected and new samples will need to be submitted after the appropriate paperwork is completed.
• NCRAD will receive a report from the IU Vector Production Facility. If the cell lines are found to be free of mycoplasma contamination, Kelley Faber at NCRAD will contact you to arrange for shipment to the repository. All arrangements for shipment of cell lines will be made by NCRAD staff and the researcher on a case-by-case basis.