Effects of Incubation Time and Method of Cell Cycle Synchronization on Collared Peccary Skin-Derived Fibroblast Cell Lines

Abstract

The success of cloning by somatic cell nuclear transfer depends on the efficiency of nuclear reprogramming, with the cycle stage of the donor cell playing a crucial role. Therefore, the aim was to evaluate three different approaches for cell cycle synchronization: (i) serum starvation (SS) for 1 to 4 days, (ii) contact inhibition (CI) for 1 to 3 days, and (iii) using cell cycle regulatory inhibitors (dimethyl sulfoxide, cycloheximide, cytochalasin B, or 6-dimethylaminopurine) for 1 and 2 days, in terms of their effects on synchronization in G0/G1 phases and viability of collared peccary skin fibroblasts. Flow cytometry analysis revealed that SS for 4 days (79.0% ± 1.6) and CI for 3 days (78.0% ± 1.4) increased the percentage of fibroblasts in G0/G1 compared to growing cells GC (68.1% ± 8.6). However, SS for 3 and 4 days reduced the viability evaluated by differential staining (81.4% ± 0.03 and 81.6% ± 0.06) compared to growing cells (GC, 95.9% ± 0.06). CI did not affect the viability at any of the analyzed time intervals. No cell cycle inhibitors promoted synchronization in G0/G1. These results indicate that CI for 3 days was the most efficient method for cell cycle synchronization in peccary fibroblasts.