Evaluating the Utility of Implementing Microfluids for Sperm Preparation Compared to Conventional Method of Density Gradient Centrifugation in a PGT-A Program: a Sibling Oocyte Study

Overview

In assisted reproductive technology (ART), sperm preparation aims to select the most viable sperm for ICSI. Unlike conventional methods like density gradients or sperm washing, microfluidic techniques mimic natural selection in the female reproductive tract by using laminar flow without centrifugation, reducing the risk of DNA damage. This method isolates highly motile sperm while filtering out debris and immotile cells. Studies show that microfluidics improve embryo quality, increase pregnancy rates, and may lead to higher euploidy rates. Additional benefits include improved safety, scalability, and shorter preparation times.

Detailed description

In assisted reproductive technology (ART), the aim of sperm preparation is to select competent spermatozoa with the highest fertilization potential to be used for insemination by intracytoplasmic sperm injection (ICSI). This makes the process of selecting sperm highly important. Several methods have been developed to mimic some of the natural selection processes that exist in the female reproductive tract. Compared to the conventional sperm preparation techniques such as density gradient or sperm wash, microfluids can select sperm by controlling fluid dynamics within millimeter diameter capillaries in two parallel laminar flow channels, mimicking what potentially sperm experiment in the female genital tract without using centrifuge which can cause DNA sperm fragmentation. Hence, this technique could select spermatozoa with increased motility since motile spermatozoa can move through the flows and be eluted separately, while the debris and immotile cells are passively transported from the entrance to the exit of the capillary canal. There is scientific evidence that for couples undergoing ICSI, the spermatozoa that were selected by using microfluids resulted in a better-quality embryo which leaded to higher pregnancy outcomes. Also, literature suggest that euploidy rates of embryos obtained using microfluids are higher that using conventional sperm sample preparation. Among the advantages that microfluidics certainly offer are, safety, scalability and reduction sperm samples preparation times.

Interventions

• Device FERTILE Plus
  The FERTILE PLUS™ method is a standardized method with an easy-to-follow protocol that is far less dependent on the skill or experience of the embryologist than other methods, such as density gradients. The FERTILE PLUS™ (850 µL) Sperm Sorting Chip is a single-use, flow-free, dual chambered, microfluidic-based sperm sorting device. FERTILE PLUS™ was previously known as Zymot, prior to a name change by the manufacturer. The lower chamber contains a sample inlet and fluid channel separated from th

Primary outcome measures

• Comparison of sperm preparation time between microfluidic and gradient methods. [Time frame: Immediately post-processing]
• Comparison of euploidy rates in embryos derived from microfluidic versus gradient-prepared sperm. [Time frame: Up to embryo biopsy (Day 5 or 6 post-fertilization)]

Secondary outcome measures (12)

• Comparison of post-processing semen parameters between microfluidic and gradient sperm preparation methods [Time frame: Immediately post-processing]
• Comparison of post-treatment semen parameters with pregnancy rates to evaluate the influence of sperm preparation methods on clinical outcomes. [Time frame: From enrollment to the end of treatment at 1 year]
• Comparison of fertilization rates between sperm processed via microfluidic and gradient methods [Time frame: Day 1 post-insemination]
• Comparison of blastulation and utilization rates of embryos derived from microfluidic vs. gradient sperm preparation [Time frame: Days 5-7 post-insemination]
• Number of Participants with Blastocyst Biopsy on Day 5, Day 6, or Day 7 by Sperm Preparation Method (Microfluidic vs. Gradient) [Time frame: Day 5 to Day 7 post-insemination]
• Comparison of blastocyst morphological quality (expansion) between sperm preparation methods [Time frame: Day 5-7 post-insemination]
• Mean Time to Key Embryo Developmental Milestones (2-Cell, 4-Cell, Blastocyst) by Sperm Preparation Method [Time frame: From fertilization to blastocyst stage (Days 0-7)]
• Comparison of post-processing semen parameters between microfluidic and gradient sperm preparation methods [Time frame: Immediately post-processing]
• Comparison of post-processing semen parameters between microfluidic and gradient sperm preparation methods [Time frame: Immediately post-processing]
• Comparison of post-processing semen parameters between microfluidic and gradient sperm preparation methods [Time frame: Immediately post-processing]
• Comparison of blastocyst morphological quality ( inner cell mass - ICM, ) between sperm preparation methods [Time frame: Day 5-7 post-insemination]
• Comparison of blastocyst trophectoderm quality (TE grades) between sperm preparation methods [Time frame: Day 5-7 post-insemination]

Eligibility criteria

Inclusion criteria

• Women with at least 8 MII per cycle after denudation (AFC≥8).
• Women of all ages.
• All embryo qualities ≥BL3CC at the time of biopsy on day 5, 6 and/or 7.
• Fresh sperm used from ejaculate with a concentration ≥1 mill/ml and ≥10% motility (A+B).
• Sperm samples with a minimum of 2 ml.

Exclusion criteria

• Frozen oocytes samples with severe oligospermia (≤1mill/ml).
• PGT-M cases
• Sperm with > 1M/ml of round cells

Criteria are shown verbatim from the registry (in English). Final eligibility is always assessed by the study center.

Healthy volunteers: Yes

Study design

Allocation

Non-randomized

Model

Parallel assignment

Masking

Open label

Primary purpose

Basic science

Study locations

United Arab Emirates · 1 center

• ART Fertility Clinics LLC — Abu Dhabi

Publications

• Chinnasamy T, Behr B, Demirci U. Microfuidic sperm sorting device for selection of functional human sperm for IUI application. Fertil Steril. 2016;105:e17-8. https://doi.org/10.1016/j.fertn stert.2015.12.063.
• Heydari A, Zabetian Targhi M, Halvaei I, Nosrati R. A novel microfluidic device with parallel channels for sperm separation using spermatozoa intrinsic behaviors. Sci Rep. 2023 Jan 21;13(1):1185. doi: 10.1038/s41598-023-28315-7. PMID 36681743
• Huang CH, Chen CH, Huang TK, Lu F, Jen Huang JY, Li BR. Design of a gradient-rheotaxis microfluidic chip for sorting of high-quality Sperm with progressive motility. iScience. 2023 Jul 17;26(8):107356. doi: 10.1016/j.isci.2023.107356. eCollection 2023 Aug 18. PMID 37559897
• Fang Y, Wu R, Lee JM, Chan LHM, Chan KYJ. Microfuidic invitro fertilization technologies: transforming the future of human reproduction. TrAC Trends Anal Chem. 2023;160:116959. https:// doi.org/10.1016/j.trac.2023.116959.
• Bastuba M, Cohen M, Bastuba A, Campbell P. Microfluidic sperm separation device dramatically lowers DFI. Fertil Steril. 2020;113(4, Supplement):E44 https://doi.org/10.1016/j.fertnstert.2020.02.096.
• Leung ETY, Lee CL, Tian X, Lam KKW, Li RHW, Ng EHY, Yeung WSB, Chiu PCN. Simulating nature in sperm selection for assisted reproduction. Nat Rev Urol. 2022 Jan;19(1):16-36. doi: 10.1038/s41585-021-00530-9. Epub 2021 Nov 5. PMID 34741158
• Quinn MM, Jalalian L, Ribeiro S, Ona K, Demirci U, Cedars MI, Rosen MP. Microfluidic sorting selects sperm for clinical use with reduced DNA damage compared to density gradient centrifugation with swim-up in split semen samples. Hum Reprod. 2018 Aug 1;33(8):1388-1393. doi: 10.1093/humrep/dey239. PMID 30007319
• Whitesides GM. The origins and the future of microfluidics. Nature. 2006 Jul 27;442(7101):368-73. doi: 10.1038/nature05058. PMID 16871203

Identifiers

NCT: NCT07093619 · 2504-ABU-011-AAb

Primary sources (government registries)

• ClinicalTrials.gov (US NIH)

View this study on ClinicalTrials.gov ↗