Cell cycle synchronization and analysis of apoptosis‐related gene in skin fibroblasts from domestic cat ( Felis silvestris catus ) and kodkod ( Leopardus guigna )

Contents The kodkod population is in constant decrease and the somatic cell nuclear transfer ( SCNT ) might help to preserve the genetic pool of this species. The cell cycle synchronization of donor cells plays a crucial role in SCNT . The objective of this research was to evaluate two different methods for quiescence induction, serum starvation ( SS ) and contact inhibition ( CI ), both for 1, 3 and 5 days, on skin fibroblast from domestic cat and kodkod. Flow cytometry analysis revealed that in domestic cat, SS and CI , both at 3 and 5 days, increased the percentage of fibroblasts in G0/G1 compared to growing cells ( GC ) ( p  < .05). In kodkod, only SS for 3 and 5 days and CI for 1 and 3 days increased the percentage of fibroblasts in G0/G1 compared to GC ( p  < .05). Viability analysis by differential staining revealed that SS for 5 days decreased the proportion of live fibroblasts in domestic cat and kodkod ( p  < .05). Regarding gene expression analysis, in domestic cat fibroblasts, no differences were found in the BAX / BCL 2 ratio in SS and CI (both at 1, 3 and 5 days) compared to GC . In kodkod fibroblasts, BAX / BCL 2 ratio was increased in CI at 3 and 5 days compared to SS at 3 and 5 days ( p  < .05). In conclusion, in kodkod fibroblasts SS for 5 days and CI after 3 days might have a negative impact on cellular viability. According to these results, we suggest SS for 3 days for cell cycle synchronization in kodkod fibroblasts.