Percoll密度梯度离心分离牛精子初探

Study on Bull Sperm Separation by Percoll Density with Centrifuge

用2.9％柠檬酸钠将奶牛原精密度调到5×10~8/ml,取其3ml经26％Percoll洗涤(1700转,25分钟),置于pH7.0、渗透压300mmol/kg的9级Percoll梯度溶液上分离(1000转,15分钟)从第9层取得精子作去Percoll处理(2500转,20分钟),再用含甘油和卵黄的牛冷冻稀释液平衡滴冻,解冻后活力可达0.4左右。分离后精子的受胎率为52.2％(47/90),母犊率为44.2％(19/43),对照组母犊率为36.8％(21/57)。说明分离后精子可正常受胎,并有提高母犊率的趋势。

Sperm concentration was adjusted to 5×108/ml by 2.9% sodium citrate solution which 3 ml was washed by 26% percoll (1700 rpm, 25 minutes), then put it in 9 degree Percoll solution with pH 7.0 and 300 mmol/kg of osmotic pressurs to centrif-Ugualizing (2500 rpm) 20 minutes. The conception rate of separated sperm after thawing was 52.2% (47/90), the rate of female calf was 44.2% (19/43) and the control group was 36.8% (21/57).