Repurposing advanced DMEM/F-12 cell culture medium to mitigate glycerol toxicity in poultry semen

= 0.044) and progressive motility increased from 19.5% to 33.2%. No differences were seen at 4°C. Membrane integrity and mitochondrial activity remained unchanged across treatments.4. The selective motility enhancement seen without mitochondrial changes suggested that adDF12 supported glycolytic rather than oxidative metabolism. Using adDF12 provided glucose (17.5 mM), pyruvate (0.5 mM) and 21 amino acids, substrates largely absent in LPC, which likely enabled flexible ATP generation under glycerol stress.5. These findings established adDF12 as a potential post-thaw resuspension medium for cryopreserved avian semen, directly supporting genetic conservation programmes.