Sperm Selection by Rheotaxis and Thermotaxis in In-Situ Handmade Microfluidics of Fluidic Walls in the Same ICSI Plate
- Conditions
- MicrofluidicsIntracytoplasmic Sperm InjectionSperm
- Interventions
- Other: Sperm selection for ICSI
- Registration Number
- NCT06243926
- Lead Sponsor
- CREA Medicina de la Reproducción SL
- Brief Summary
Microfluidics technologies are proving their capability to select suitable sperm for ICSI, especially those that take advantage of the rheotaxic properties of sperm migration. In contrast to other sperm preparation methods such as Wash-Swim-Up and Density- Gradients-Centrifugation (DGC), microfluidics disregards washing and centrifugation steps along the procedure. However, microfluidics devices are costly and likely to fail to select motile sperm in dispermic samples. The investigators have recently described a novel approach for sperm selection by In-Situ handmade rheotaxis microfluidics of fluidic walls in the same ICSI plate. This methodology integrates a swim-over based on horizontal migration with a positive rheotaxis zone. The present study aims to deliver an unbiased comparison between two sperm selection techniques: DGC and In-Situ handmade rheotaxis microfluidics of fluidic walls (isM).
- Detailed Description
Microfluidics technologies stand as the latest sperm selection methodology for ICSI. Increasingly, publications show novel and ingenious microfluidics strategies to integrate sperm biomimicry during in vitro sperm selection while simplifying the IVF workflow. Microfluidics are time-efficient methods which reduce the risks associated with handling, gamete mix-up and ROS production. However, microfluidics devices are costly and likely to fail to select motile sperm in dispermic samples. Aiming to outline the application of microfluidics in ART, the investigators conceived a lab-on-a-chip approach for sperm selection by In-Situ rheotaxis handmade microfluidics of fluidic walls. This microfluidics system allows selecting sperm for ICSI in the same ICSI-dish. Thus, from a microvolume of the raw semen sample, only using microfluidics, disregarding centrifugation, washing, plasticware, other cells, products or materials, and using the same culture medium needed to prepare the rest of the ICSI plate.
A previous proof-of-concept study showed that this methodology efficiently selected suitable sperm for ICSI. The investigators also demonstrated that the microfluidic protocol effectively separates at least 20 progressive spermatozoa in less than 15 minutes in a clean microdroplet, free of any remaining seminal plasma. A subsequent non-inferiority study showed that the microfluidic method performs as well as density gradient centrifugation to achieve ICSI outcomes such as fertilization and day-5 blastocyst formation rate, proving that In-Situ handmade rheotaxis microfluidics of fluidic walls are also effective in supporting pre-implantation embryonic development in vitro.
Recruitment & Eligibility
- Status
- RECRUITING
- Sex
- All
- Target Recruitment
- 50
- Patients undergoing ICSI
- Female age under 40 years old
- Number of mature oocytes (MII) upon oocyte retrieval ≥ 6
- Male age under 50 years old
- Total number of progressive sperm ≥ 1 million
- Patients with surgically retrieved sperm for ICSI
- Cases using cryopreserved oocytes
- Cases using cryopreserved sperm
Study & Design
- Study Type
- OBSERVATIONAL
- Study Design
- Not specified
- Arm && Interventions
Group Intervention Description In-Situ rheotaxis handmade microfluidics (isM) Sperm selection for ICSI The isM is a novel protocol to select sperm for ICSI based on microfluidics and sperm guidance responses to rheotaxis and thermotaxis. The isM protocol proved its efficacy and effectivity for ICSI in previous proof of concept and non-inferiority studies, respectively. Density gradient centrifugation (DGC) Sperm selection for ICSI DGC is a protocol to select sperm based on their density and isopycnic points. It is a conventional protocol commonly used in IVF laboratories.
- Primary Outcome Measures
Name Time Method Fertilization Rate (FR) 16 to 20 hours post-ICSI (day 1) The proportion of two-pronuclei zygote divided by the total number of matured and microinjected oocytes (2pn/MII, %)
Day 5 usable blastocyst rate (D5UBR) 108 to 113 hours post-ICSI (day 5) The proportion of embryos classified as blastocysts at day 5 of development, which has been transferred and/or cryopreserved.
Blastocyst quality grade 108 to 113 hours post-ICSI (day 5) Blastocysts are categorized following the scale described by the Spanish Association of Reproduction Biology Studies (ASEBIR). The ASEBIR classification categorizes blastocysts based on their morphology into four groups: from A (highest, better outcome) to D (lowest, worse outcome).
- Secondary Outcome Measures
Name Time Method Morphokinetics profile (MK PNa) from 16 to 20 hours post-ICSI (day 1) The proportion of embryos showing the appearance of pronuclei (PNa) out of the expected time frame.
Morphokinetics profile (MK PNf) from 24 to 25 hours post-ICSI (day 1) The proportion of embryos showing the fading of pronuclei (PNf) out of the expected time frame.
Morphokinetics profile (MK tSB) from 99 to 101 hours post-ICSI (day 5) The proportion of embryos starting to blastulate (tSB) out of the expected time frame.
Morphokinetics profile (MK t4) from 37 to 41hours post-ICSI (day 2) The proportion of embryos showing 4 cells (t4) out of the expected time frame.
Morphokinetics profile (MK tB) from 108 to 113 hours post-ICSI (day 5) The proportion of embryos starting to blastulate (tB) out of the expected time frame.
Morphokinetics profile (MK t8) from 51 to 69 hours post-ICSI (day 3) The proportion of embryos showing 8 cells (t8) out of the expected time frame.
Morphokinetics profile (MK t2) from 24 to 28 hours post-ICSI (day 1) The proportion of embryos showing 2 cells (t2) out of the expected time frame.
Morphokinetics profile (MK tM) from 81 to 96 hours post-ICSI (day 4) The proportion of embryos with full cell compaction (morula; tM) out of the expected time frame.
Implantation Rate (IR) 5 to 6 gestational weeks The proportion of embryos that were transferred that develop to a gestational sac
Ongoing clinical pregnancy Rate (OCP) 6 to 8 gestational weeks The proportion of embryos that were transferred that develop at least to the stage of fetal heart activity
Trial Locations
- Locations (1)
CREA Medicina de la Reproducción
🇪🇸Valencia, Spain