Exploring the effects of roscovitine, serum starvation, and contact inhibition at G0/G1 arrest in northern tiger cat dermal fibroblasts.

Abstract

Nuclear reprogramming studies are important tools in conserving wild felids, with efficacy depending on efficient G₀/G₁ cell cycle arrest methodologies. This study evaluated different culture conditions at G₀/G₁ arrest and the quality of northern tiger cat fibroblasts. Cells from four animals were assigned to groups: 7.5 and 15 µM roscovitine (RSV) for 24 and 48 h; serum starvation (SS) for 24, 48, 72, and 96 h; and contact inhibition (CI) for 24, 48, and 72 h. Cells with 50-60% confluence were used as control. The cell quality parameters included morphology, and viability and apoptotic levels were assessed through microscopic analysis, while cell cycle phases were evaluated using flow cytometry. RSV affected the cell viable percentage and morphology with the increase of concentration and exposure time. Moreover, RSV did not improve the cells at G₀/G₁. CI did not significantly affect cell quality or increase the proportion of cells in G₀/G₁ phase. Interestingly, SS for 24 h increased the cells at G₀/G₁. However, SS affected the apoptosis levels. The SS for 24 h is the most efficient method of G₀/G₁ arrest for northern tiger cat fibroblasts. However, adjustments are still necessary to optimize cell arrest for northern tiger cat fibroblasts.