scholarly journals Elimination of apoptotic spermatozoa by magnetic-activated cell sorting improves the fertilization rate of couples treated with ICSI procedure

Andrology ◽  
2013 ◽  
Vol 1 (6) ◽  
pp. 845-849 ◽  
Author(s):  
A. Sheikhi ◽  
M. Jalali ◽  
M. Gholamian ◽  
A. Jafarzadeh ◽  
S. Jannati ◽  
...  
Keyword(s):  
Cell Sorting ◽  
Fertilization Rate ◽  
Magnetic Activated Cell Sorting

P–081 Microfluidic sorting does not improve clinical outcomes compared to magnetic activated cell sorting (MACS) in Assisted Reproduction

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
C González-Ravina ◽  
A Pachec. Castro ◽  
M Cru. Palomino ◽  
A Requen. Miranda
Keyword(s):  
Clinical Outcomes ◽  
Cell Sorting ◽  
Fertilization Rate ◽  
Male Factor ◽  
Ovum Donation ◽  
Sperm Dna Damage ◽  
Sperm Dna ◽  
Frozen Embryos ◽  
Microfluidic Sorting ◽  
Magnetic Activated Cell Sorting

Abstract Study question Does microfluidic sorting improve clinical outcomes over magnetic activated cell sorting (MACS) in ovum donation cycles? Summary answer Performing microfluidic sorting does not seem to improve clinical outcomes compared to MACS in ovum donation cycles. What is known already Novel sperm selection techniques, such as magnetic activated cell sorting (MACS), have been described as useful procedures to increase reproductive outcomes when male factor is present. Because of centrifugation steps associated to swim-up or density-gradient can induce sperm DNA fragmentation, microfluidic sperm sorters are being used to isolate motile human spermatozoa based on fluid dynamics and avoiding sample manipulation. This new technology has been shown to reduce the level of sperm DNA damage, especially double strand breaks, but an improvement of clinical outcome by using this technique remain unclear. Study design, size, duration Prospective and observational study to evaluate the efficacy of a sperm sorting technique based on microfluidic technology versus a technique based on the removal of apoptotic spermatozoa by MACS. The study was performed between May 2019 to January 2021 in IVI Madrid and IVI Sevilla. All men attending for an ovum donation cycle during the aforementioned study period were included. The exclusion criteria were sperm concentration <5 mill/mL and <15% of progressive motile spermatozoa. Participants/materials, setting, methods Seminal samples from couples participating in the study were divided into two aliquots; each of them was processed according to one of the study methods. Subsequently, each of the processed sperm samples was used to microinject half of the oocytes obtained during oocyte retrieval. In all case, a single-embryo transfer was performed. Variables were expressed as mean values and standard deviations. Statistical analysis was performed by ANOVA and Chi-squared where applicable; significance established under 0.05. Main results and the role of chance We included 48 couples in the study; of these, n = 31 transferred an embryo derived from a MACS processed sperm sample, while n = 17 received an embryo from the microfluidic one. Groups were homogeneous in terms of number of transferred embryos and frozen embryos, usable blastocyst rate and fertilization rate; results were as follows for MACS and microfluidic processing respectively: number of transferred embryos (1.0±0.0 vs. 1.0±0.0, p = 0.978); number of frozen embryos (1.6±0.5 vs. 1.4±0.7 p = 0.168); usable blastocyst rate (40.7% vs. 43.1%, p = 0.384); and fertilization rate (80.4% vs. 75.3%, p = 0.075). However, according clinical outcomes, we observed significant differences in implantation rate (74.2% vs. 58.8%, p < 0.001) and in clinical pregnancy rate (74.2% vs. 58.8%, p < 0.001); finally, the miscarriage rate was similar between the two groups of study (6.4% vs. 5.8%, p = 0.876). Limitations, reasons for caution This study has not considered the indication in male factor couples due to a high degree of double-strand DNA fragmentation. Therefore, more specific studies are required to determine in which patients, microfluidics sorter selection would significantly improve clinical outcomes. Wider implications of the findings: In an unselected population, magnetic activated cell sorting significantly improves clinical outcomes compared to a microfluidic technique, so this latter method should not be recommended without a male factor indication associated with sperm DNA damage. The proposed microfluidic technology does not seem to offer a flow-free approach to select spermatozoa. Trial registration number NCT04061486

P–235 Are morphokinetics parameters and profitability of an assisted reproduction cycle related to sperm selection technique? Microfluidic sperm sorter chip versus magnetic activated cell sorting

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
D Agud. Garcillan ◽  
E Santamarí. López ◽  
C Gonzále. Ravina ◽  
A Pachec. Castro ◽  
M Cru. Palomino ◽  
...  
Keyword(s):  
Assisted Reproduction ◽  
Cell Sorting ◽  
Statistical Significance ◽  
Fertilization Rate ◽  
Sperm Selection ◽  
Fragmented Dna ◽  
Significant Difference ◽  
Blastocyst Rate ◽  
Sperm Sorting ◽  
Magnetic Activated Cell Sorting

Abstract Study question Does microfluidic sperm sorter offer any biological improvement over magnetic activated cell sorting (MACS)? Summary answer Microfluidic approach for selecting high profile spermatozoa is as good as magnetic activated cell sorting in terms of morphokinetic, fertilization and good quality blastocyst rate. What is known already Microfluidic sperm sorter chip is a method for select non-fragmented DNA sperm. We know that the use of these non-fragmented DNA sperms can improve the clinical results and the useful blastocyst rate. As several studies have shown previously, embryos morphokinetics parameters are affected by culture medium, ovarian stimulation, oxygen tension, origin of the oocytes, or the age of the patient, this is why we wanted to compare if the cleavage times using time lapse technology, are different depending on the sperm selection method used to select sperm with the non-fragmented DNA. Study design, size, duration Prospective and observational study performed between May 2019 to January 2021 in IVI Madrid and IVI Sevilla. Seminal samples from couples participating in the study were divided into two aliquots; each of them was processed according to one of the study methods. 53 couples were included in the study. Half of the oocyte from each donor were microinjected with sperm selected through MACS (n = 281) and the other half through a microfluidic device (n = 275). Participants/materials, setting, methods These oocytes were microinjected with both types of sperm samples and incubated in EmbryoScope. Cellular events studied in this study included cellular divisions until blastocyst stage, appearance and fading of some cellular structures and the duration of the first, second and third cellular cycle (cc1, cc2 and cc3) as well as their synchrony (S1, S2, S3). Data were exported from the EmbryoViewer data base. We perform an ANOVA statistical analysis to analyze the data. Main results and the role of chance No significant differences between both sperm selection methods were found regarding the time of cell division (from T2 to Tblstocyst), the cellular cycles duration (cc1, cc2 and cc3) or the synchrony of the cellular cycles divisions (s1, s2 and s3). However, a clear trend towards statistical significance has been found in both duration of cc2 (p = 0.052) being longer in MACS embryos than in microfluidic sperm sorting embryos, and in the expansion of the blastocyst, which occurs earlier in embryos that come from MACS than in those that come from microfluidic sperm sorting (p = 0.097). These two events could indicate a better embryo cleavage dynamic in the case of MACS embryos, with a better blastocyst expandability and the necessary time to carry out all the biological events that must occur in the cc2. However, significant difference was found in the direct cleavage from 1 to 3 cells embryo stage, which is one of the adverse events that more affects embryo implantation, being higher in microfluidic sperm sorting group (p = 0,037). Finally, the fertilization rate (73.1% vs 76.9%) and the good quality blastocyst rate (53.7% vs 56.5%) were higher in MACS embryos than in microfluidic sperm sorting embryos, although no significant differences were found. Limitations, reasons for caution This study has been performed in donated oocytes, so these results may not be extrapolated to other groups of assisted reproduction patients. However, more data are needed to draw firm conclusions. Furthermore, it´s crucial to increase the sample size to check if the trends founded reach statistical significance. Wider implications of the findings: Microfluidic sorting of unprocessed semen in un unselected population is as efficacious as magnetic activated cell sorting according embryo morphokinetic, fertilization rate and useful blastocyst rate. Microfluidic sperm sorting does not show clinical advantage over MACS considering this data collection. Trial registration number NCT04061484

scholarly journals Production of Female Bovine Embryos with Sperm Sorted by Magnetic Activated Cell Sorting

2018 ◽  
Vol 63 (1) ◽  
pp. 45-52
Author(s):  
Lili ZHUANG ◽  
Ju Lan CHUN ◽  
Junbo WU ◽  
Ji Hye LEE ◽  
Eun Young KIM ◽  
...  
Keyword(s):  
Cell Sorting ◽  
Bovine Embryos ◽  
Magnetic Activated Cell Sorting

scholarly journals Expression and functional analyses of ephrin type-A receptor 2 in mouse spermatogonial stem cells†

2019 ◽  
Vol 102 (1) ◽  
pp. 220-232 ◽  
Author(s):  
Hiroko Morimoto ◽  
Mito Kanatsu-Shinohara ◽  
Kyle E Orwig ◽  
Takashi Shinohara
Keyword(s):  
Stem Cells ◽  
Cell Sorting ◽  
Type A ◽  
Spermatogonial Stem Cells ◽  
Fibroblast Growth Factor 2 ◽  
Self Renewal ◽  
Spermatogonial Transplantation ◽  
Short Hairpin ◽  
Functional Analyses ◽  
Magnetic Activated Cell Sorting

Abstract Spermatogonial stem cells (SSCs) undergo continuous self-renewal division in response to self-renewal factors. The present study identified ephrin type-A receptor 2 (EPHA2) on mouse SSCs and showed that supplementation of glial cell-derived neurotrophic factor (GDNF) and fibroblast growth factor 2 (FGF2), which are both SSC self-renewal factors, induced EPHA2 expression in cultured SSCs. Spermatogonial transplantation combined with magnetic-activated cell sorting or fluorescence-activated cell sorting also revealed that EPHA2 was expressed in SSCs. Additionally, ret proto-oncogene (RET) phosphorylation levels decreased following the knockdown (KD) of Epha2 expression via short hairpin ribonucleic acid (RNA). Although the present immunoprecipitation experiments did not reveal an association between RET with EPHA2, RET interacted with FGFR2. The Epha2 KD decreased the proliferation of cultured SSCs and inhibited the binding of cultured SSCs to laminin-coated plates. The Epha2 KD also significantly reduced the colonization of testis cells by spermatogonial transplantation. EPHA2 was also expressed in human GDNF family receptor alpha 1-positive spermatogonia. The present results indicate that SSCs express EPHA2 and suggest that it is a critical modifier of self-renewal signals in SSCs.

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