血清饥饿法同步化牛卵巢颗粒细胞周期的研究

摘要

研究了体细胞制备过程中细胞培养时间、血清饥饿浓度和饥饿对间对体细胞周期同步化的影响.体细胞为牛卵巢颗粒细胞,细胞用碘化丙啶(PI)染色法,用流式细胞仪检测细胞周期.细胞传代培养至第2、10和25代,在0.5％和0.05％的血清饥饿浓度下诱导处理2、3、4和5d.试验结果表明,第2、10和25代的细胞在G0/G1期的比例没有差异(P＞0.05).培养液中添加0.5％ FCS和0.05％ FCS较对照组(10％FCS)提高了细胞G0/G1期的比例(P＜0.05),0.5％FCS和0.05％ FCS试验组获得了相近的细胞G0/G1期比例(P＞0.05).血清饥饿至第5天的细胞较第2天的细胞有高的G0/G1期的比例(P＜0.05).结果显示,体外培养至不同代的牛卵巢颗粒细胞对G0/G1期的比例没有影响,血清饥饿和延长血清饥饿的时间能够有效诱导体细胞进入G0/G1期.%The effects of culture time, serum starvation concentration and starvation time on cell cycle synchronization of in vitro cultured bovine granulosa cells were investigated by cell cycle analysis with flow cytometer. Bovine granulosa cells were cultured to P2, P10 and P25, and then cultured in 0.5% and 0.05% FCS serum starvation for 2, 3, 4 and 5 d. The results showed that bovine granulosa cells of P2, P10 and P25 had no significant differences in the percentages of G0/G1 phase(P> 0.05). The percentages of G0/G1 phase in 0.5% and 0.05% FCS treat group were significantly higher than that in control group (10% FCS} (P<0.05), and there were no significant differences between the treat groups (P>0,05), The percentage of G0/G1 phase of cells cultured for 5d was significantly higher than the cells cultured for 2 d. It indicated that the cultured passages of bovine granulosa cells in vitro had no effect on the percentages of G0/G1 phase, serum starvation and prolonging serum starvation time could effectively induce the cells into G0/G1 phase.