通过密度梯度离心法制备的人类精子中衰老减少且核DNA完整性得以保留。
Reduced senescence and retained nuclear DNA integrity in human spermatozoa prepared by density gradient centrifugation.
作者信息
Morrell J M, Moffatt O, Sakkas D, Manicardi G C, Bizzaro D, Tomlinson M, Nilsson H, Holmes P V
机构信息
Nidacon International, Gothenburg, Sweden.
出版信息
J Assist Reprod Genet. 2004 Jun;21(6):217-22. doi: 10.1023/b:jarg.0000040237.47026.0f.
Abstract
PURPOSE
To investigate whether removal of extraneous cells and immotile spermatozoa from a sperm preparation by density gradient centrifugation could help to maintain normal spermatozoa in a viable state and retain their deoxyribonucleic acid integrity.
METHODS
Sperm motility was assessed on a daily basis in aliquots of neat semen, extended semen, and spermatozoa prepared on a PureSperm density gradient. At the same time, aliquots of each sperm sample were preserved for TUNEL assay and nick translation.
RESULTS
Spermatozoa prepared using density gradient centrifugation survived three times as long as spermatozoa in neat semen or in extended semen. Both deoxyribonucleic acid integrity and sperm motility were retained in the gradient preparations.
CONCLUSIONS
Preparing spermatozoa by density gradient centrifugation is advantageous in prolonging sperm survival and maintaining deoxyribonucleic acid integrity, presumably by removing sources of reactive oxygen species. Stored spermatozoa could be used for a second attempt at fertilization if oocyte immaturity was suspected.
摘要
目的
研究通过密度梯度离心从精子制剂中去除多余细胞和不活动精子是否有助于使正常精子保持存活状态并保留其脱氧核糖核酸完整性。
方法
每天对纯精液、稀释精液以及在PureSperm密度梯度上制备的精子的等分试样进行精子活力评估。同时，保存每个精子样本的等分试样用于TUNEL检测和缺口平移。
结果
使用密度梯度离心制备的精子存活时间是纯精液或稀释精液中精子的三倍。梯度制剂中同时保留了脱氧核糖核酸完整性和精子活力。
结论
通过密度梯度离心制备精子有利于延长精子存活时间并保持脱氧核糖核酸完整性，推测这是通过去除活性氧来源实现的。如果怀疑卵母细胞不成熟，储存的精子可用于第二次受精尝试。
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