<abstract xmlns="http://www.w3.org/1999/xhtml">

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.
</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.

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

Universidade Federal Rural do Semi-Árido

Annals of Animal Science

This work is licensed under the Creative Commons Attribution 4.0 International License.

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...

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

Published Online: Aug 05, 2021

Page range: 925 - 938

Received: Jun 17, 2020

Accepted: Sep 16, 2020

DOI: https://doi.org/10.2478/aoas-2020-0103

Keywords
G/G phase, karyoplast, somatic cell nuclear transfer, wild mammals

© 2021 Alana Azevedo Borges et al., published by Sciendo

This work is licensed under the Creative Commons Attribution 4.0 International License.

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

Published Online: Aug 05, 2021

Page range: 925 - 938

Received: Jun 17, 2020

Accepted: Sep 16, 2020

DOI: https://doi.org/10.2478/aoas-2020-0103

KeywordsG/G phase, karyoplast, somatic cell nuclear transfer, wild mammals

© 2021 Alana Azevedo Borges et al., published by Sciendo

This work is licensed under the Creative Commons Attribution 4.0 International License.

Keywords
G/G phase, karyoplast, somatic cell nuclear transfer, wild mammals