Sperm enrichment from poor semen samples by double density gradient centrifugation in combination with swim-up for IVF cycles

Abstract

Sperm preparation in iVf cycles using density gradient centrifugation (DGc) in combination with swim-up (SU) has been widely adopted in reproductive centres worldwide. it is a fact that the sperm recovery rate following one DGc from poor semen samples (showing liquefaction defects/containing too many unresolvable clots or rare sperm) is relatively low. our results showed that double DGc (DDGC) is effective at increasing the sperm recovery rate from poor semen samples. However, DDGC may increase the mechanical stress of sperm, thereby potentially impairing embryo development. Therefore, it is necessary to evaluate the safety of using sperm prepared by DDGC/SU for IVF cycles. In this study, we retrospectively analysed the data generated from a total of 529 IVF cycles (from June 2017 to June 2018), and these IVF cycles contributed 622 transfer cycles (from June 2017 to December 2018) in Changzhou Maternal and Child Health Care Hospital. Of them, 306 IVF cycles and the related 355 transfer cycles (normal semen samples prepared by DGC/SU) were set as the normal group, while 223 IVF cycles and the related 267 transfer cycles (poor semen prepared by DDGC/SU) were set as the observation group. The main outcome measures, including the normal fertilization rate, top D3 embryo formation rate, blastocyte formation rate, clinical pregnancy rate and live birth rate, birth weight and duration of pregnancy, were compared between the two groups. Compared to semen in the DGC/SU group, semen in the DDGC/SU group showed increased levels of the DNA fragmentation index (DFI) and reduced sperm concentration, percentage of progressive motility (PR) sperm, and percentage of normal morphology sperm. The indicators reflecting in vitro embryo development and clinical outcomes were similar in the DGC/SU group and DDGC/SU group, including the normal fertilization rate, top D3 embryo formation rate, blastocyte formation rate, pregnancy rate, implantation rate, spontaneous abortion rate, live birth rate, birth weight and duration of pregnancy. Furthermore, we found that the 1PN zygote formation rate was significantly lower in the DDGC/SU group than that in the DGC/SU group. We concluded that oocytes fertilized by sperm from poor semen samples separated by DDGC/SU achieved the same outcomes as oocytes fertilized by sperm from normal semen separated by DGC/SU, suggesting that DDGC/SU is an effective and safe method of sperm enrichment for poor semen samples in IVF. The main contribution of the present study is the verification of the effectiveness of DDGC/SU in improving sperm recovery from poor semen samples and the safety of using sperm prepared by DDGc/ SU for iVf.

AMH has no role in predicting oocyte quality in women with advanced age undergoing IVF/ICSI cycles

“…After oocytes retrieval, insemination was performed by the conventional IVF or ICSI method in IVF-plus medium (Vitrolife, Sweden). The observation of pronuclei, embryo culture and score were performed as we described previously 24 . Briefly, 18 h after insemination, zygotic embryos (day 1) with 2 pronuclei were considered normal fertilization and were placed into G1 plus medium (Vitrolife, Sweden) for further culture.…”

“…Briefly, 18 h after insemination, zygotic embryos (day 1) with 2 pronuclei were considered normal fertilization and were placed into G1 plus medium (Vitrolife, Sweden) for further culture. On day 3, the morphology of the embryos was evaluated and scored according to a previously described method 24 . Grade I and II embryos were taken as top embryos, while Grade III embryos were taken as non-top embryos.…”

Differential impact of four sperm preparation techniques on sperm motility, morphology, DNA fragmentation, acrosome status, oxidative stress, and mitochondrial activity: A prospective study

“…Sperm preparation techniques have previously been evaluated without reaching a firm consensus. 19,[21][22][23][24][25][26][27][28][29] Several reasons could explain the discrepancies between the published reports such as the number and types of techniques being compared (eg, density gradient, semen and/or pellet swim-up, double density gradient or density gradient followed by swim-up), the included participants (eg, fertile and/ or infertile men), the sample size, and the measured outcomes (eg, sperm DNA fragmentation, embryo development or pregnancy rates).…”

“…[15][16][17] Previous studies found that SARS, 1 of the 3 epidemic coronaviruses to emerge in the past 20 years and that shows similar clinical presentations to COVID-19, 18 could cause orchitis 19 and focal testicular atrophy. 20 Considering the tens of millions of COVID-19 cases and that men are more vulnerable to COVID-19 than women, [21][22][23] it is imperative to determine the effect of COVID-19 on male reproduction. 24 Several studies have been performed on this topic.…”

“…20 Debate on the differential impact of these procedures on motile sperm characteristics is still ongoing due to differences in the published reports regarding the study design, the sperm selection techniques included and the outcomes measured. 19,[21][22][23][24][25][26][27][28][29] A Cochrane review conducted on six randomized clinical trials (RCTs) indicated low-quality evidence to support the superiority of one sperm preparation technique over others (wash and centrifugation; swim-up; density gradient) in terms of live birth in IUI treatment. The authors highlighted the limitations of the analyzed RCTs such as risk of bias (eg, blinding, allocation concealment) and the dissimilarities in the primary outcome assessed.…”