磁珠分选法与密度梯度离心法分离精子对辅助生殖结局的影响

Magnetic-activated cell sorting （MACS） versus density gradient centrifugation （DGC） for the selection of human sperm in assisted reproductive techniques

目的比较新型磁珠分选法（MACS-DGC）与传统的密度梯度离心法（DGC）分离精子对辅助生殖（ART）结局的影响。方法回顾分析2014年1至12月在浙江大学医学院附属妇产科医院生殖中心进行精子DNA碎片率（DFI）检测并进入体外受精-胚胎移植（IVF-ET）周期的病例。根据DFI将病例分为3组：DFI异常（DFI≥120）且使用新型精子磁珠分选法（A组）；DFI异常（DFI≥120）且使用传统精子密度梯度离心法（B组）；DFI正常（DFI〈20）且使用传统精子密度梯度离心法（C组）。根据受精方式的不同，将常规体外受精（IVF）标记为1组，卵细胞质内单精子显微注射（ICSD标记为2组。对各组的正常受精率、优胚率、临床妊娠率，胚胎种植率、抱婴率等临床结果进行统计分析。结果在IVF三组中（A1、B1、C1）正常受精（2PN）率、优胚率、临床妊娠率，胚胎种植率、抱婴率差异均无统计学意义（均P〉0．05）。在高DFI的ICSI患者（A2、B2）中，A2组对比B2组发现优胚率（87．8％比46．3％，P〈0．001）、临床妊娠率（66．7％比29．2％，P=0．031）、胚胎种植率（50．0％比19．5％，P=0．010）、抱婴率（66．7％比25．0％，P=0．016）差异均有统计学意义。结论新型精子磁珠分选法对比传统梯度分离法对于IVF的临床结果差异无统计学意义，但在ICSI受精周期中临床结局得到了显著改善。

Objective To investigate whether the assisted reproductive techniques （ART） outcomes can be improved by using magnetic-activated cell sorting combined isolate density gradient eentrifugation （MACS-DGC）, and to compare the results with the conventional isolate density gradient centrifugation （DGC） method. Methods In the period from January to December 2014, patients who attended the andrology laboratory for sperm DNA fragmentation analysis and underwent the in vitro fertilization and embryo transfer （IVF-ET） treatment were divided into 3 groups, and each group has two subgroups according to the way of fertilization ：in vitro fertilization （IVF） as subgroup 1 and intracytoplasmic sperm injection （ICSI） as subgroup 2. Group A （ A1, A2 ） ： DNA fragmentation index （DFI） ≥ 20 and using MACs; Group B （B1, B2） ： DFI≥20 and using isolate method; Group C （C1, C2） ： DFI 〈20 and using isolate method. The fertilization rate, good-quality embryos, implantation rate, pregnancy rate and live-birth rate of each group were analyzed. Results Similar results were obtained among groups for all the parameters compared： On IVF cycles no significant difference was observed in fertilization rate, percentage of good-quality embryos on day 3, implantation rate, positive beta-hCG tests, and live-birth rate among the three subgroups （A1 vs B1 vs C1 ）. Between the subgroups of ICSI cycles （A2 vs B2 vs C2） the percentage of high-quality embryos, pregnancy rate, implantation rate and live-birth rate were all significantly higher in the MACS-DGC （A2） group compared to DGC alone （B2）. Conclusion Based on the results, MACS as a part of sperm preparation technique will not improve sperm fertilizing potential, embryos quality, implantation rate, pregnancy rate or live-birth rate of IVF, but it may help select the most fertile sperm and to improve clinical outcomes of ICSI.