Effects of Incubation Time and Method of Cell Cycle Synchronization on Collared Peccary Skin-Derived Fibroblast Cell Lines
Annals of Animal Science(2021)
摘要
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 G(0)/G(1) 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 G(0)/G(1) 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 G(0)/G(1). These results indicate that CI for 3 days was the most efficient method for cell cycle synchronization in peccary fibroblasts.
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关键词
G(0)/G(1) phase,karyoplast,somatic cell nuclear transfer,wild mammals
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