梯度密度离心中离心力、时间和梯度液体积对精子回收率和受精潜能的影响

The effects of centrifugal force, time and gradient liquid volume of gradient density centrifugation gradient on the recovery of sperm and fertilization potential

目的探究梯度密度离心中离心力、时间和梯度液体积对前向运动精子回收率和受精潜能的影响。方法实验分新鲜精液组和冷冻精液组各30例。每例精液充分混匀后,平均分为4份,进行下列各组实验:(1)组1为80%和40%的梯度离心液各0.5m L,300×g,离心20min;(2)组2为80%和40%的梯度离心液各1.0m L,300×g,离心20min;(3)组3为80%和40%的梯度离心液各1.0m L,1 200×g,离心5min;(4)组4为80%和40%的梯度离心液各0.5m L,1 200×g,离心5min。分别检测处理前后每组精液浓度、前向运动精子百分率、正常形态率以及精子DNA碎片率,以分析离心力、时间和梯度液体积对精子前向运动精子回收率和受精潜能的影响。结果无论是新鲜精液,还是冷冻精液,组1、组3和组4前向运动精子的回收率和精子的正常形态率以及精子的DNA碎片率与组2比较无明显差异(P>0.05)。结论将现行的300×g,离心20min的离心方法,改为1 200×g,离心5min,梯度离心液体积从1.0mL降低至0.5mL,并不显著影响精子正常形态率、前向运动精子的回收率和精子的受精潜能,是可行的。

Objective To investigate the effect of centrifugal force, time and gradient liquid volume of the gradient density centrifugation on the recovery of the progressive motile sperm and fertilization potential. Methods Semen samples were divided into 30 cases of fresh semen group and 30 cases of frozen semen group. And each sample was divided into 4 subgroups： Subgroup 1： 80% and 40% gradient liquid volume each 0.5 m L, 300×g centrifugation for 20 minutes. Subgroup 2： 80% and 40% gradient liquid volume each 1m L, 300×g centrifugation for 20 minutes. Subgroup 3： 80% and 40% gradient liquid volume each 1m L, 1 200×g centrifugation for 5 minutes. Subgroup 4： 80% and 40% gradient liquid volume each 0.5m L, 1 200×g centrifugation for 5 minutes. Recovery of the sperm and fertilization potential, sperm concentration, progressive motile sperm, normal morphology and sperm DNA fragmentation rates were detected before and after treatment in each group. Results There were no significant differences in the normal morphology of sperm, the recovery rate of the progressive motile sperm and the rate of sperm DNA fragments between subgroup 2 and other subgroups（P 〉 0.05）,regardless of fresh or frozen semen sample. Conclusion Although 300 g centrifugation 20 min of centrifugation was changed into 1 200×g, centrifuged for 5 minutes, and 1.0m L gradient liquid volume was changed into 0.5 m L, sperm normal morphology rate, progressive motile sperm rate and sperm fertilization potential remained unchanged