Magnetic cell sorting of semen containing spermatozoa with high DNA fragmentation in ICSI cycles decreases miscarriage rate

Sánchez-Martín, Pascual; Dorado-Silva, Mónica; Sánchez-Martín, Fernando; Martínez, Mercedes González; Johnston, Stephen D.; Gosálvez, Jaime

doi:10.1016/j.rbmo.2017.01.015

Cited by 23 publications

(19 citation statements)

References 35 publications

(41 reference statements)

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“…In a sub‐analysis, the in‐house designed swim‐out methods were compared to the commercial Magnetic‐activated cell sorting test (MACS ® ART Annexin V, Miltenyi Biotec, 2016). Several studies have shown the potential of MACS selected spermatozoa for fertility treatment (Martínez et al., 2018; Sánchez‐Martín et al., 2017). These studies assumed an association of sperm apoptosis with sperm DNA damage (Martínez et al., 2018; Sánchez‐Martín et al., 2017), and therefore, SDF is often used as outcome parameter in studies comparing semen preparation methods with MACS ® .…”

Section: Discussionmentioning

confidence: 99%

“…Several studies have shown the potential of MACS selected spermatozoa for fertility treatment (Martínez et al., 2018; Sánchez‐Martín et al., 2017). These studies assumed an association of sperm apoptosis with sperm DNA damage (Martínez et al., 2018; Sánchez‐Martín et al., 2017), and therefore, SDF is often used as outcome parameter in studies comparing semen preparation methods with MACS ® . The mean SDF rate was significantly reduced after density gradient and MACS ® (1.69 ± 3.57%) compared with density gradient prepared spermatozoa (2.46 ± 7.70%) or native spermatozoa (7.01 ± 4.31%) ( p ≤ .001).…”

Section: Discussionmentioning

confidence: 99%

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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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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

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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“…They suggested that not all species are suitable for MACS separation method. In our knowledge, most of the research for MACS separation or the combination of DGC and MACS used human samples (de Vantéry Arrighi, Lucas, Chardonnens, & De Agostini, 2009;Gil, Shalom, Sivira, Carreras, & Checa, 2013;Martin et al, 2017;Romany et al, 2017). DGC has been proven to effectively remove nonviable spermatozoa.…”

Section: Discussionmentioning

confidence: 99%

The Effect of Density Gradient Centrifugation and Magnetic-Activated Cell Sorting with the Combination of Both on Bovine Spermatozoa Kinematics, Viability and Acrosome Status

Osman

Bakri

Jaffar

et al. 2017

JAS

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Conventional and novel spermatozoa separation techniques have their own strength and weaknesses. Both are widely used in assisted reproductive technologies (ARTs). In this study, combinations of two separation methods against single separation methods were evaluated. Freshly ejaculated semen was collected and divided into five groups; control, density gradient centrifugation (DGC), magnetic activated cell sorting (MACS), DGC-MACS and MACS-DGC; based on the sequence of separation. Evaluation of spermatozoa kinematics, viability and acrosome status for each group were done by computer assisted semen analysis (CASA), hypo-osmotic swelling test (HOST) and chlorotetracycline (CTC) staining respectively. Results show that DGC yield the fastest spermatozoa velocity and highest percentage of viable spermatozoa fraction compared to other groups. However, they also produced a high percentage of acrosome-reacted spermatozoa. Spermatozoa isolated through DGC-MACS shows the highest percentage of non-capacitated spermatozoa. However, spermatozoa isolated through DGC-MACS yields slower spermatozoa velocity population and lower percentage of viable sperm compared to spermatozoa isolated through DGC. Isolation of spermatozoa through MACS-DGC yields slower population of spermatozoa velocity with a lower percentage of non-capacitated spermatozoa compared to spermatozoa isolated through DGC-MACS. DGC separation and its combination with MACS had yield different spermatozoa quality. This distinct characteristic is likely suitable for in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) procedure respectively.

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“…In a previous study, we tested one of these techniques (magnetic‐activated cell sorting of non‐apoptotic spermatozoa; MACS) and observed that it might improve the quality of selected spermatozoa and the outcome of ICSI cycles 24 . Several other studies have also shown the benefits of using MACS for IVF/ICSI 25‐27 . To analyze the possible benefits of such spermatozoa selection methods on the level of DNA methylation, we performed reduced representation bisulfite sequencing (RRBS) to determine the genome‐wide methylation profiles of our samples.…”

Section: Introductionmentioning

confidence: 99%

Minor DNA methylation changes are observed in spermatozoa prepared using different protocols

Štimpfel

Vrtačnik-Bokal

2020

Andrology

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Background DNA methylation patterns can show transgenerational inheritance and are influenced by lifestyle and environmental factors. It is suggested that these patterns can be changed by assisted reproductive technology. Objectives To evaluate the impact of two different sperm preparation methods, conventional density gradient centrifugation (DGC) vs. density gradient centrifugation followed by magnetic‐activated cell sorting (MACS) of non‐apoptotic spermatozoa, on sperm DNA methylation profile. Materials and methods We analyzed semen of patients included in our IVF treatment program. Half of the semen from each included patient was prepared for ICSI using the DGC method and the other half with DGC followed by MACS. The remaining samples were processed for DNA methylation analysis with reduced representation bisulfite sequencing (RRBS). In addition to the DNA methylation profile, we assessed the morphology and DNA fragmentation of spermatozoa. Results RRBS analysis revealed that the average genome‐wide methylation level was similar between both groups (DGC vs. MACS group) and ranged from 0.53 to 0.56. Furthermore, RRBS analysis identified 99 differentially methylated regions (DMRs) and 800 differentially methylated positions (DMPs). In the DGC group, 43 DMRs and 392 DMPs were hypermethylated whereas 56 DMRs and 408 DMPs were hypomethylated compared with those in the MACS group. When DMRs and DMPs were annotated to genes, 3 genes associated with imprinting were found: IGF2, PRDM16, and CLF4/BRUNOL4. The percentage of morphologically normal spermatozoa (MACS vs. DGC; 14.0 ± 10.8 vs. 13.2 ± 10.0; P = .335) and of spermatozoa with fragmented DNA of patients with RRBS analysis (22.9 ± 21.1% vs. 34.4 ± 21.2; P = .529) were also similar between groups. Discussion and Conclusion Although the average genome‐wide level of sperm DNA methylation was similar in both sample groups, a distinctive number of methylation changes were observed in DMR and DMP levels. A larger number of samples should be analyzed and additional sperm preparation methods should be tested to confirm our findings.

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Magnetic cell sorting of semen containing spermatozoa with high DNA fragmentation in ICSI cycles decreases miscarriage rate

Cited by 23 publications

References 35 publications

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

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

The Effect of Density Gradient Centrifugation and Magnetic-Activated Cell Sorting with the Combination of Both on Bovine Spermatozoa Kinematics, Viability and Acrosome Status

Minor DNA methylation changes are observed in spermatozoa prepared using different protocols

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