Comparison of the effects between optimization of semen with the floating plate and density gradient centrifugation with swim-up in split semen samples
Objective:To optimize human semen with floating plate and density gradient centrifugation combined with upstream method in order to clarify the optimization effects and compare the difference of sperm quality between the two methods.Methods:Eighty-five normal semen samples and 59 oligoasthenozoospermia semen samples were obtained,and each sample was divided into the unprocessed semen group(1 ml),density-gradient centrifugation and swim-up group(DGU group,1 ml)as well as floating plate group(FP group,1 ml).The unprocessed semen group did not undergo any treatment after semen liquefaction,the DGU group was treated with density-gradient centrifugation with swim-up,while the FP group was treated with a microfluidic device called floating plate method.The sperm concentration,forward motility sperm percentage,and normal morphology sperm percentage were compared among the three subgroups of the unprocessed semen group,DGU group and FP group.Sperm mitochondrial membrane potential and sperm DNA fragmentation index were detected with flow cytometry,and sperm reactive oxygen species levels were evaluated with a single sperm reactive oxygen species analyzer,and the electron microscopy was used for the observation of sperm ultrastructure.The different indicators were compared among three groups.Results:Compared with the unprocessed semen group,the percentage of progressive motile sperm,normal morphology sperm,and mitochondrial membrane potential of sperm in the DGU group and FP group were significantly increased(P＜0.05),while the DNA fragmentation index of sperm was significantly reduced(P＜0.05)for both normal semen and oligoasthenozoospermia semen.There were no significant differences in these indicators between the FP and DGU groups.For normal semen,the sperm concentration of FP group was significantly higher than that of DGU group(P=0.029),and the level of reactive oxygen species in single sperm was significantly lower than that in the unprocessed semen group(P=0.038).For the oligoasthenozoospermia semen,there were no significant differences in the levels of reactive oxygen species of single sperm among the three subgroups(P＞0.05).Electron microscopy observation revealed that there were no significant abnormal sperm in the normal semen of three subgroups.There were abnormal cell membrane breakage and mitochondrial swelling in the sperm of the DGU group,but no obvious abnormality was observed in the sperm of the FP group for the oligoasthenozoospermia semen.Conclusions:DGU and FP methods can effectively retrieve sperm with better progressive motility,and better morphology as well as function.The FP method can obtain sperm with less reactive oxygen species and less damage,and it is easier to operate than the DGU.
Density gradient centrifugationFloating plate methodSingle sperm reactive oxygen speciesSperm DNA fragmentation indexSperm mitochondrial membrane potential
万方数据
维普
