目的:目前可用于在胞浆内单精子注射(ICSI)之前利用透明质酸(HA)结合进行精子选择的商业产品被广泛使用,但具有一些缺点。为了潜在地规避这些限制,我们将使用自制透明质酸(smHA)试剂的ICSI与使用SpermSlow的ICSI进行了比较。
方法:使用密度梯度离心和向上游动程序(N=10/组)分离的精子,对试剂与塑料或玻璃底盘上精子的结合进行定量。此外,我们调查了ICSI前使用的HA试剂与HA-ICSI辅助生殖后的临床结局之间的关系(N=81).
结果:smHA试剂表现出与人精子极其稳定的结合。在塑料和玻璃皿上,smHA试剂中精子的结合时间明显长于SpermSlow(塑料:60.0±0.0minvs.2.7±5.9min,P<0.001;玻璃:60.0±0.0minvs.2.5±1.8min,P<0.001)。用smHA试剂的HA-ICSI(128/160,80.0%)和用SpermSlow的HA-ICSI(171/231,74.0%,P=0.184)。使用smHA试剂(74/101,73.3%)的HA-ICSI衍生的受精卵的胚泡发育频率明显高于SpermSlow(76/131,58.0%,P=0.019)。生化妊娠率,临床妊娠,胎心运动,活产,使用smHA试剂的HA-ICSI和使用SpermSlow的HA-ICSI之间的流产没有显着差异。
结论:与SpermSlow相比,使用smHA试剂的HA-ICSI的囊胚形成率更高。临床结果,不包括囊胚形成,HA-ICSI后使用smHA试剂和使用SpermSlow相同。精子与smHA试剂的结合在60分钟的时间过程中没有减弱。总之,该试剂可以缩短和简化HA-ICSI程序,因为smHA可以与任何培养皿材料一起使用,更容易观察纺锤体或评估胞浆内形态。
OBJECTIVE: Commercial products currently available for sperm selection utilizing hyaluronic acid (HA)
binding prior to intracytoplasmic sperm injection (ICSI) are widely used but have some disadvantages. To potentially circumvent these limitations, we compared ICSI using a self-made hyaluronic acid (smHA) reagent with ICSI using SpermSlow.
METHODS: The
binding of the reagents to spermatozoa on plastic- or glass-bottom dishes was quantitated using spermatozoa that were isolated by density-gradient centrifugation and swim-up procedures (N = 10/group). Additionally, we investigated the relationship between the HA reagent used prior to ICSI and clinical outcomes after assisted reproduction with HA-ICSI (N = 81).
RESULTS: The smHA reagent exhibited extremely stable
binding to human spermatozoa. The
binding time of spermatozoa was significantly longer in the smHA reagent than in SpermSlow on both plastic and glass dishes (plastic: 60.0 ± 0.0 min vs. 2.7 ± 5.9 min, P < 0.001; glass: 60.0 ± 0.0 min vs. 2.5 ± 1.8 min, P < 0.001). There were no significant differences in the normal fertilization rate between HA-ICSI with the smHA reagent (128/160, 80.0%) and HA-ICSI with SpermSlow (171/231, 74.0%, P = 0.184). The frequency of the blastocyst development from the HA-ICSI-derived zygote was significantly higher with the smHA reagent (74/101, 73.3%) than with SpermSlow (76/131, 58.0%, P = 0.019). The rates of biochemical pregnancy, clinical pregnancy, fetal heart movement, live birth, and miscarriage were not significantly different between HA-ICSI with the smHA reagent and HA-ICSI with SpermSlow.
CONCLUSIONS: The blastulation rate was higher for HA-ICSI with the smHA reagent as compared with SpermSlow. Clinical outcomes, excluding blastulation, after HA-ICSI were the same using smHA reagent and using SpermSlow. Spermatozoa
binding to the smHA reagent was not attenuated over a 60-min time course. In conclusion, this reagent may shorten and simplify HA-ICSI procedures because smHA can be used with any dish material, making it easier to observe the spindle or assess intracytoplasmic morphology.