Mycoplasma testing is a required part of release testing to ensure a cell product is sterile and safe to infuse into patients. To meet this safety standard with an increasing product volume, our group aimed to enable batching of DNA isolation and Mycoplasma PCR testing of cultured cell products to conserve reagents and decrease technologist time. Towards this aim, we evaluated the stability of Mycoplasma DNA in snap-frozen cells in media stored at -80°C compared to the supplier-recommended isolation from freshly harvested cells. DNA from three cultured cell products - CAR T-cells, TGFβ-imprinted NK cells, and NK cells - was isolated both fresh on the day of harvest and after storage at -80°C following snap-freezing. Cultured cells in media were spiked with three different Mycoplasma strains at 10CFU/mL prior to DNA isolation: M. fermentans, M. orale, & M. pneumoniae. These samples were run in tandem with DNA isolated from unspiked cells in media. Mycoplasma PCR testing was performed using the MycoTOOL Mycoplasma Real-Time PCR Kit. All extraction, inhibition, and PCR controls passed acceptance criteria. All spiked cells in media frozen at -80°C for 14 (n=2) or 15 days (n=1) tested positive for the Mycoplasma species in 8 of 8 PCR reaction wells. All unspiked cells in media samples tested negative. In summary, Mycoplasma DNA is stable in snap-frozen cultured cells in media stored at -80°C up to 15 days. This validation allows sample batching for PCR testing to reduce technologist time and maximize reagent utilization. Our group plans to perform additional testing of spiked, snap-frozen cells in media stored for > 15 days at -80°C, the stability of spiked DNA stored for > 3 days at -20°C, and to validate increased Proteinase K volume for DNA isolation to overcome varying protein content in distinct culture media.