P>培养时间和传代次数对山羊输卵管上皮细胞周期和凋亡的影响/P>

目的 利用山羊输卵管上皮细胞(OECs)探讨体外培养时间和传代次数对细胞生长的影响.方法 建立山羊OECs的体外培养体系,用流式细胞术分别对半数贴壁、刚贴满(贴壁1d)、贴满3d和贴满5d,以及原代、Ⅲ代和Ⅴ代细胞分别进行细胞周期和凋亡检测.结果细胞贴满3d和传至Ⅲ代时,细胞凋亡比率较低,处于静止期(G0/G1)细胞明显降低,极性线粒体受损明显降低;培养时间延长和传代次数增加后,凋亡比率显著升高,处于增殖期(S,G2/M)的比率显著降低,极性线粒体受损明显升高.结论采用OECs培养胚胎时,细胞应控制在Ⅲ代以内,且贴满时间不超过3d.

Effect of days after confluence and passages on the cell cycle and apoptosis of goat oviductal epithelial cells

Objective The experiment was conducted with the objective of determining effects BR> of culture time and subculture on the growth of goat oviductal epithelial cells (OECs). Methods We firstly succeeded in culturing goat OECsEM>in vitro/EM>, and then the cell cycle was examined by flow cytometric analysis and cell apoptosis was determined by Annexin-V and PI staining in goat OECs at different days of confluence and different passages at day 1 of confluence. Results BR>The percentages of （GSUB>0/SUB>/GSUB>1/SUB>） stage, late apoptotic + necrotic and cells with reduced number of polarized mitochondria decreased when OECs were subcultured within Ⅲ passage and confluent less than three days. However, the percentages of S + GSUB>2/SUB>/M stage decreased but late apoptotic + necrotic and cells with reduced number of　polarized mitochondria increased with prolonged culture and passage. Conclusion A better development of co cultured embryos w