A new micro swim-up procedure for sperm preparation in ICSI treatments: preliminary microbiological testing

Palini, Simone; Primiterra, Mariangela; Stefani, Stefania; Pedna, Maria Federica; Sparacino, Monica; Farabegoli, P.; Benedetti, Serena; Bulletti, Carlo; Sambri, Vittorio

doi:10.5935/1518-0557.20160023

Cited by 9 publications

(10 citation statements)

References 27 publications

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“…Semen is not sterile [2] and may contain microorganisms even after processing for ART. Although most of the microorganisms detected in semen samples are non-pathogenic commensals or contaminants, their presence has a great significance on in vitro fertilization (IVF), a treatment in which the natural defense of the female genital tract is largely bypassed [3]. Therefore, different approaches have been proposed to reduce the microbial contamination and load in IVF culture media by improving semen preparation and embryo culture protocols.…”

Section: Introductionmentioning

confidence: 99%

The complex microbiome from native semen to embryo culture environment in human in vitro fertilization procedure

Štšepetova

Baranova

Simm

et al. 2020

Reprod Biol Endocrinol

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Background: Only a few microbial studies have conducted in IVF (in vitro fertilization), showing the high-variety bacterial contamination of IVF culture media to cause damage to or even loss of cultured oocytes and embryos. We aimed to determine the prevalence and counts of bacteria in IVF samples, and to associate them with clinical outcome. Methods: The studied samples from 50 infertile couples included: raw (n = 48), processed (n = 49) and incubated (n = 50) sperm samples, and IVF culture media (n = 50). The full microbiome was analyzed by 454 pyrosequencing and quantitative analysis by real-time quantitative PCR. Descriptive statistics, t-, Mann-Whitney tests and Spearman's correlation were used for comparison of studied groups. Results: The study involved normozoospermic men. Normal vaginal microbiota was present in 72.0% of female partners, while intermediate microbiota and bacterial vaginosis were diagnosed in 12.0 and 16.0%, respectively. The decreasing bacterial loads were found in raw (35.5%), processed (12.0%) and sperm samples used for oocyte insemination (4.0%), and in 8.0% of IVF culture media. The most abundant genera of bacteria in native semen and IVF culture media were Lactobacillus, while in other samples Alphaproteobacteria prevailed. Staphylococcus sp. was found only in semen from patients with inflammation. Phylum Bacteroidetes was in negative correlation with sperm motility and Alphaproteobacteria with high-quality IVF embryos. Conclusion: Our study demonstrates that IVF does not occur in a sterile environment. The prevalent bacteria include classes Bacilli in raw semen and IVF culture media, Clostridia in processed and Bacteroidia in sperm samples used for insemination. The presence of Staphylococcus sp. and Alphaproteobacteria associated with clinical outcomes, like sperm and embryo quality.

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Section: Introductionmentioning

confidence: 99%

The complex microbiome from native semen to embryo culture environment in human in vitro fertilization procedure

Štšepetova

Baranova

Simm

et al. 2020

Reprod Biol Endocrinol

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“…On the contrary, the novel procedure here reported, requires a total of 10 min to prepare the sperm, with a time saving of about 50 min compared with the direct swim-up procedure and further 75-80 min when including the density gradient step in the preparation. On the other hand, this procedure is cheaper for avoiding the cost of media that need to be necessarily used with the classical technique of sperm preparation while it has also been demonstrated in the absence of bacterial contaminations [28,29].…”

Section: Discussionmentioning

confidence: 99%

A fast and safe technique for sperm preparation in ICSI treatments within a randomized controlled trial (RCT)

Baldini

Silvestris

et al. 2020

Reprod Biol Endocrinol

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Recently a novel method based on horizontal sperm migration in injection dishes has been introduced as an additional tool for preparation of semen sample in assisted reproductive technology (ART) procedures. In the present study, we evaluated both timing and reproductive outcomes in a randomized controlled study including 1034 intra-cytoplasmic sperm injection (ICSI) procedures followed by fresh embryo transfer. Couples enrolled were divided into two subgroups , namely conventional swim-up method (Group A), and horizontal sperm migration in injection dishes (Group B). No significant differences were found between groups with respect to fertilization rate, implantation success, clinical pregnancy outcomes and ongoing pregnancies. On the contrary, both cleavage and blastocyst rates were statistically higher in Group B, suggesting superior efficiency and safety of this innovative technique also including time-saving and cheaper costs as compared to the classical swim-up sperm preparation. Our data support the interpretation of the horizontal sperm migration as a promising procedure for semen preparation in ART cycles.

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“…To our knowledge only the groups of Palini et al. (2016) and De Martin et al. (2017), presented a non‐commercialised modification of the classic swim‐up procedure, called the micro swim‐up and the positive rheotaxis extended droplet (PRED), respectively, directly correlated with the selection of spermatozoa for ICSI.…”

Section: Introductionmentioning

confidence: 99%

A methodological validation of an easy one‐step swimout semen preparation procedure for selecting DNA fragmentation‐free spermatozoa for ICSI

et al. 2020

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The main purpose of this methodological paper was to describe a recently designed one‐step ICSI semen preparation swim‐out method (called swim‐ICSI) and to compare its efficacy with our conventional two‐step swim‐out method for the selection of motile spermatozoa for ICSI with minimal DNA damage. In this observational cohort study, 42 fresh ejaculate sperm samples for ICSI were included to compare the new swim‐ICSI with the conventional swim‐out. In a sub‐analysis (n = 20), both in‐house designed ICSI preparation methods were compared with a commercial magnetic‐activated cell sorting test (MACS®). Sperm DNA fragmentation (SDF), using Halosperm®, was determined at different time points during sperm preparation: on the native sample (a), after density gradient centrifugation (DG) (b), on the motile (A + B) spermatozoa selected with conventional swim‐out post‐DG (c) and selected with swim‐ICSI method post‐DG (d). For a subgroup (n = 20), SDF was also calculated after MACS (e). The mean SDF significantly reduced after EACH preparation step and reduced to almost zero in the recovered A + B spermatozoa when the semen prepared with DG was further processed for ICSI (swim‐ICSI vs. swim‐out, p = .001). In conclusion, the optimised one‐step and fine‐tuned swim‐ICSI technique shows the possibility to select a population of spermatozoa with almost zero SDF to be used in ICSI treatments.

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A new micro swim-up procedure for sperm preparation in ICSI treatments: preliminary microbiological testing

Cited by 9 publications

References 27 publications

The complex microbiome from native semen to embryo culture environment in human in vitro fertilization procedure

The complex microbiome from native semen to embryo culture environment in human in vitro fertilization procedure

A fast and safe technique for sperm preparation in ICSI treatments within a randomized controlled trial (RCT)

A methodological validation of an easy one‐step swimout semen preparation procedure for selecting DNA fragmentation‐free spermatozoa for ICSI

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