多级梯度离心后X/Y精子浓度分析

罗熙; 李永刚; 李明颖; 曾鹏; 晏家骢

多级梯度离心后X/Y精子浓度分析

昆明理工大学附属云南省第一人民医院生殖医学科

基金项目:

基金：云南省卫生科技计划基金资助项目 (2012WS0073);

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出版历程
- 收稿日期: 2017-03-16

Analysis of X/Y Sperm Concentration after Multistage Gradient Centrifugation

Funds:

基金：云南省卫生科技计划基金资助项目 (2012WS0073);

摘要

摘要: 目的在辅助生殖中, 利用多层密度梯度离心的方法来分选X/Y精子, 以检验常规IVF-ET中, X/Y精子比例是否失衡.方法使用Sperm Grad TM (Vitrolife, Sweden) 与Sperm Rinse TM (Vitrolife, Sweden) 配制10层梯度液, 分别为100%、95%、90%、85%、80%、75%、70%、65%、60%、55%.在离心管中制成多层梯度后, 每层梯度间分界明显.选择液化正常、浓度约50×106/mL、活动率约50%, 畸形率正常的5人精液混合后, 去除个体差异取3 mL进行梯度离心.分别收集每层梯度底部精子, 利用双色X/Y FISH探针 (Vysis) 进行FISH检测, 统计X/Y精子比例.结果 100%、95%、90%、85%、80%五层梯度中X精子含量比例分别为:51.41%、51.41%、52.19%、50.99%、54.10%.结论每层梯度液中X/Y精子含量比例虽有不同, 但差异并不明显.现在辅助生殖技术中, 利用商品化精子梯度分离液, 即使使用多层梯度进行离心分离, 也不会造成性别比例失衡.
- 梯度离心法 /
- X/Y精子分选 /
- FISH /
- 荧光原位杂交
Abstract: Objective In assisted reproduction, the X/Y sperm was sorted by multi density gradient centrifugation to examine whether the proportion of X/Y sperm was unbalanced in conventional IVF-ET.Method The use of Sperm Grad (Vitrolife, Sweden) and Sperm Rinse (Vitrolife, Sweden) to prepare 10 gradient liquid layer, were 100%, 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%.After the multilayer gradient was made in the centrifuge tube, the boundary between each gradient was obvious.Choose liquefaction normal, concentration of about 50×106/mL, activity rate of about 50%, low deformity rate of 5 people, after mixing semen, removing individual differences, and take 3 mL mixture gradient centrifugation.Collect each gradient bottom sperm, using two-color X/Y FISH probe (Vysis) for detection, and statistics X/Y sperm percentage.Results 100%, 95%, 90%, 85%, 80%, The proportion of X/Y sperm in each gradient was: 51.41%, 51.41%, 52.19%, 50.99%, 54.10%.Conclusions Although the proportion of X/Y sperm in each gradient was different, the difference was not obvious.In assisted reproductive technology, the use of commercialized sperm washing fluid, even if the use of multi-layer gradient centrifugation, will not cause gender imbalance.
- Gradient centrifugation /
- X/Y sperm sorting /
- FISH /
- Fluorescence in situ hybridization

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参考文献(16)

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