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.

中文翻译：

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孵育时间和细胞周期同步化方法对领兽皮皮肤衍生成纤维细胞系的影响

通过体细胞核转移克隆的成功取决于核重编程的效率，供体细胞的循环阶段起着至关重要的作用。因此，目的是评估三种不同的细胞周期同步方法：（i）血清饥饿（SS）1至4天；（ii）接触抑制（CI）1至3天；以及（iii）使用细胞周期调节抑制剂（二甲基亚砜，环己酰亚胺，细胞松弛素B或6-二甲基氨基嘌呤）持续1天和2天，这取决于它们对G ₀ / G ₁相同步性的影响以及猫齿状皮肤皮肤成纤维细胞的活力。流式细胞仪分析显示，SS 4天（79.0％±1.6）和CI 3天（78.0％±1.4）会增加成纤维细胞在G ₀ / G中的百分比与生长细胞GC相比为₁（68.1％±8.6）。然而，与生长中的细胞（GC，95.9％±0.06）相比，SS处理3天和4天会降低通过差异染色评估的生存力（81.4％±0.03和81.6％±0.06）。CI在任何分析的时间间隔内均不影响生存力。没有细胞周期抑制剂促进G ₀ / G _1中的同步。这些结果表明，CI 3天是最重要的方法，用于同步化成纤维细胞的细胞周期同步。