Tissue Culture Shared Resource

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Preparation of Conditioned Media for Mycoplasma Detection

Mycoplasma Testing

Mycoplasma tests should be performed on all cells arriving to Lombardi from outside laboratories and as a routine check on existing stocks.

All cell lines used in TCSR shared labs must be demonstrated to be mycoplasma negative.

The TCSR provides incubators and hoods free of charge for quarantining new cell lines prior to testing.

Sample Preparation

  1. Cell lines must be cultured for at least two passages prior to testing.
  2. Cell lines must be cultured in the absence of antibiotics and selection agents during this time (such as Pen-Strep, G418, gentamycin, geneticin, fungizone, puromycin, or zeocin). Mycoplasma is reduced but not eliminated in the presence of routine tissue culture antibiotics, i.e., you will get a false negative!
  3. Media should be collected after it has been in contact with cells for three days.
  4. Transfer approximately 10 ml of the media into a centrifuge tube.
  5. Bring the sample and a completed request form to TCSR.
  6. Please store samples at 4oC until you bring them to the TCSR.
  7. Label your sample.