外源bcl—2基因和脂质体对鸡卵泡颗粒细胞体外分裂和凋亡的影响

选用正在产蛋的罗曼鸡，取其排序第3的卵泡分离层颗粒层细胞，用含血清培养基培养，并用含脂质体每孔0．0，0．6，1．2，2．4，4．8μL和bcl-2基因重组质粒每孔0．2μg转染卵泡颗粒细胞。用流式细胞术等方法分析转基因后的源代细胞和传代细胞的生长与凋亡情况。结果发现，与未转染组相比，转染后的传代细胞分裂速度较快（P＜0．01）、凋亡比率较低，G2／M＋S期细胞极显著高于对照（P＜0．01），且与脂质体浓度有剂量依赖关系。在转染时间为12h、DNA含量为每孔0．2μg的条件下，脂质体最佳转染浓度为每孔1.2μL。这些结果进一步证明，外源bcl-2基因或脂质体具有促进细胞分裂、抑制细胞凋亡、延长细胞寿命的作用。

Effect of bcl-2 gene andliposome on division and apoptosis of ovarian granular cells in laying hens

Granulosa cells were isolated from the follicles of each laying Romen hens and incubated in 2 mL of medium 199 supplementing with 5% calf serum for 3 days,and then were transfected by bcl-2 gene using 0.0，0.6，1.2，2.4 and 4.8 μL of liposome in 24-well plates.The growth and apoptosis of primary cells and subcultured cells were investigated by flow cytometry methods.The results showed that bcl-2 gene transfected cells grew more quickly (P＜0.01)，with lower ratio of apoptosis and higher ratio of cells at G2/M＋S phase(P＜0.01)in liposome dose dependence,in comparison with the control during the subculture period.The optimal concentration of liposome was 1.2 μL per well under the condition of 12 hours of transfection and 0.2 μg per well of bcl-2 re-combined plasmid.These results further indicated that bcl-2 gene or liposome play important roles in control of growth,development and apoptosis of ovarian granular cells in hens.