血清饥饿、汇合培养及放线菌酮对奶牛成纤维细胞周期的影响

在动物克隆研究中，研究者普遍认为位于细胞周期的G0+G1期的二倍体细胞对于核移植中供核细胞的重新程序化是必需的。本文探讨了血清饥饿、汇合培养及放线菌酮（CHX）处理对不同传代次数的体外培养奶牛成纤维细胞周期分布的影响。流式细胞仪分析结果显示：第3代和第13代细胞经血清饥饿处理72h后，细胞周期分布与对照差异显著(P<0.05)；汇合培养可以显著增加奶牛成纤维细胞处于G0+G1期细胞数。CHX处理第3代细胞经CHX处理后处于G0+G1期的细胞数差异不显著，而第13代细胞处理后差异显著。结果表明体外培养奶牛成纤维细胞高代对血清饥饿、汇合培养及CHX处理更敏感。

Effects of Serum Deprivation, Confluence and Cycloheximide on Cell Cycle of Dairy Cow Fibroblasts

In animal cloning, it is generally believed that the inactive diploid G0 or G1 stage of the cell cycle is beneficial to initiate cell-cycle coordination and reprogramming following transfer of the donor nucleus. In order to provide more G0+G1 stage cells for dairy cattle cloning, we carried out a flow cytometric analysis of the cell cycle of fibroblasts from the ear of dairy cow at different passage numbers after different periods of serum, after full confluence, after cycloheximide（CHX） treatment. Serum starvation effectively increased the percentage of G0+G1 sage cells after 72h. Significantly higher percentage of cells in G0+G1 was apparent when dairy cattle fibroblast cells cultured in confluence. No significant difference were observed on the 3rd passage cultures exposed to CHX, however, on the 13th passage an increase on proportion of cells was observed after all periods of exposure(p<0.05). This date indicates that high passage cells in vitro are more susceptible to serum starvation, confluence cultured and CHX G0+G1 synchronization.