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 ◽  
...  

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

2013 ◽  
Vol 30 (4) ◽  
pp. 479-485 ◽  
Author(s):  
Monica Gil ◽  
Valerie Sar-Shalom ◽  
Yolisid Melendez Sivira ◽  
Ramon Carreras ◽  
Miguel Angel Checa

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
M Saleh. Novin ◽  
Z Zandie ◽  
M Bakhtiari ◽  
R Aflatoonian

Abstract Study question What effect does Annexin-V MACS method have on sperm selection by pLCZ1 expression and blastocyst rate in male DNA fragmentation? Summary answer AnnexinV-MACS method could be a good choice for sperm selection with high pLCZ1 expression and high blastocyst rate in high DNA fragmentation male factor. What is known already Sperm selection based on morphology and motility in ART techniques, is not enough for choosing the best sperm especially in male factor patients. In Annexin-V magnetic activated cell sorting (MACS) technique, apoptotic sperm are separated from non-apoptotic one by negative selection. So, this method can improved quality of compaction rate in embryo. PLCζ is oocyte activating factors that it starts oscillations of calcium ca2+ in oocyte and it has a significant effect on fertilization and implantation. Study design, size, duration Semen samples from 30 male factor infertile couples with high DFI (DFI>30%) were selected and divided into two group (control and experimental) in each patient. Participants/materials, setting, methods Control was washed with DGC and experimental one was selected by MACS-DGC. Retrieved eggs in each patient, were divided in 2. Both group were injected by DGC and MACS respectively. Semen parameters and DFI (SCD test) were analyzed before and after processing. After ICSI, rate of fertilization, embryo development and blastocyst formation were evaluated. Real time PCR evaluate expression of PLCζ. Comparison between results of two groups was assessed by SPSS analysis. Main results and the role of chance Results showed that, sperm motility and morphology after MACS method (45%, 1.7%) was significantly higher than DGC method (40%, 1.1%) and before washing (35%, 0.9%). Percent of DFI in MACS group (36%) was significantly decreased compared to DGC (45%) and primitive group (55%). The number of oocytes were injected in DGC group was 93 and in MACS group was 111. Fertilization rate in both groups was almost the same (72.07% in MACS vs 73.11 in DGC). Rate of day 3 embryo with good grade in MACS group (72. 5%) was significantly higher than DGC (51.47%) (P < 0.05). The blastocyst rate in MACS-DGC group (69.69%) was significantly increased compared to DGC group (48%). PLCζ gene expression in MACS-DGC was significantly higher than DGC group (p-value=0.046). Limitations, reasons for caution This experiment was performed invivo. Wider implications of the findings: Sperm selection by MACS-DGC method can improve sperm motility, morphology and reduce sperm DFI. No significant difference was observed in fertilization rate, but percent of high-quality embryo on days 3 and 5 was significantly higher by this method,also it can be suggested as a good choice for patients with highDF. Trial registration number Not applicable


Andrology ◽  
2013 ◽  
Vol 1 (6) ◽  
pp. 845-849 ◽  
Author(s):  
A. Sheikhi ◽  
M. Jalali ◽  
M. Gholamian ◽  
A. Jafarzadeh ◽  
S. Jannati ◽  
...  

2020 ◽  
Vol 9 (12) ◽  
pp. 3976
Author(s):  
Alberto Pacheco ◽  
Arancha Blanco ◽  
Fernando Bronet ◽  
María Cruz ◽  
Jaime García-Fernández ◽  
...  

Magnetic-activated cell sorting (MACS) can be used to separate apoptotic sperm with high proportions of fragmented DNA from the rest, thus improving the overall quality of the seminal sample. Therefore, the aim of this retrospective study was to investigate the efficiency of the MACS technique to increase reproductive outcomes in patients with high levels of sperm DNA fragmentation (SDF) undergoing intracytoplasmic sperm-injection (ICSI) cycles. In this study, we analyzed a total of 724 assisted-reproduction-technique (ART) cycles that were divided into two groups: the study group (n = 366) in which the MACS selection technique was performed after density-gradient centrifugation (DGC), and the control group (n = 358) in which only DGC was used for sperm selection. Reproductive outcomes were analyzed in both groups according to three different ART procedures: preimplantation genetic testing for aneuploidy (PGT-A), and autologous and oocyte-donation cycles. The MACS group showed significantly lower miscarriage rates in autologous ICSI cycles, higher pregnancy rates in oocyte-donation cycles, and a significant increase in live-birth rates in both autologous and oocyte-donation cycles. Overall, these results suggested that the MACS technique can be effectively used to eliminate sperm with high SDF levels, and therefore may help to improve reproductive outcomes in couples undergoing ART.


2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
C González-Ravina ◽  
A Pachec. Castro ◽  
M Cru. Palomino ◽  
A Requen. Miranda

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


2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
J Y Lo ◽  
S T Tee

Abstract Study question Does Hyaluronic acid(HA) binding sperm selection prior ICSI produce better outcome compare to PVP? Summary answer No significant difference in fertilization rate, blastocyst utilization rate, pregnancy rate and miscarriage rate in HA vs PVP group. What is known already HA is natural, non-sulphated glycosaminoglycan secretion that abundantly found in the cervical mucus and the cumulus oophorus complex (COC). In in-vivo, HA binding sites on the sperm plasma membrane indicate sperm maturity, mature sperm reaching HA-coated COC can bind and digest HA and subsequent hyperactivation further facilitate fertilization. In IVF, the use of HA tries to mimic in-vivo to select mature sperm with high DNA integrity prior to ICSI. In HA, movement of mature sperm is ‘slowed’ thus allow the selection of sperm to be used in ICSI. Sperm immaturity is known to associated with aneuploidy incidence in the embryos. Study design, size, duration ICSI cycle using HA (N = 83) was adapted from January to December 2020 while ICSI cycle using PVP (N = 133) was adapted from January 2018 to December 2019. Mean age of patient were 35.64±4.33 vs 34.15±4.75 for HA vs PVP group respectively. Fertilization rate, blastocyst utilization rate, pregnancy rate and miscarriage rate were recorded. This study included all ICSI cycle and both frozen and fresh embryo transfer data. Surgical retrieved sperm was excluded from this study. Participants/materials, setting, methods A 1.5ul of treated spermatozoa suspension was connected with a pipette tip to a 5ul droplet of fresh holding medium (Cook Gamete Buffer). Simultaneously, a 5ul droplet of HA medium (Origio SpermSlow) was connected to the 5ulL droplet of holding medium in a ICSI dish (Sparmed Oosafe) covered with oil (Vitrolife Ovoil). Sperm which were ‘slowed’ in the Sperm slow droplets with normal morphology according to WHO 2010 guideline were selected for ICSI at 400X. Main results and the role of chance The fertilization rate of the HA vs PVP- binding sperm are 68.6% vs 66.2%. As for the blastocyst utilisation rate is 61.6% vs 73.22% for HA vs PVP- binding sperm group. Pregnancy and miscarriage rate for HA vs PVP are 42.3% vs 51.5% and 19.4% vs 26.2% respectively. There was no significant difference in the fertilization rates, blastocysts utilisation rate, pregnancy rate and miscarriage rate between the HA vs PVP- binding sperm groups (P < 0.05). However, a trend of higher pregnancy (51.5% vs 42.3%; P = 0.279) and miscarriage rate were observed in the PVP group (26% vs 19%; P = 0.545) as compared to the HA group, but the difference was not statistically significant. The reason behind this might be the HA assist to select the mature sperm with higher DNA integrity and low frequency of chromosomal aneuploidies which contribute to the lower miscarriage rate in the study. Limitations, reasons for caution This is a retrospective study on HA binding sperm selection vs PVP prior to ICSI. Further research which includes a large number of RCT sample size should be warranted. Wider implications of the findings: The HA- sperm binding selection ICSI might only be beneficial to certain group of patients (high- DNA fragmentation sperm). A larger RCT study may be necessary to establish a relationship between HA-sperm binding selection vs aneuploidy rate via PGT analysis. Trial registration number Not applicable


2021 ◽  
Vol 116 (3) ◽  
pp. e156
Author(s):  
Maria Gil Julia ◽  
Irene Hervas ◽  
Ana Navarro-Gomezlechon ◽  
Laura Mossetti ◽  
Fernando Quintana ◽  
...  

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