Microfluidic Chip Method Versus Density-gradient Centrifugation Method in IVF

A Randomized Double-blind Controlled Study of the Cumulative Live Birth Rate of IVF Following Sperm Preparation by Microfluidic Chip Method Versus Density-gradient Centrifugation Method

Infertile patients attending IVF treatment at the Centre of Assisted Reproduction and Embryology, Queen Mary Hospital and Kwong Wah Hospital will be recruited during ovarian stimulation for IVF. Subsequently, they will be randomly assigned on the day of oocyte retrieval by a laboratory staff into one of the following two groups: (1) the microfluidic chip group and (2) the density gradient group for sperm preparation and subsequent use in fertilization. Other IVF procedures will be the same as our usual practice. Both patients and clinicians were blinded from the group allocation i.e. a double blind study. The primary outcome is the cumulative live birth rate defined as the number of pregnancies leading to live birth within 6 months of randomisation.

Study Overview

• Status: Recruiting
• Conditions: Infertility
• Intervention / Treatment: Device: Microfluidic chip; Device: Density-gradient centrifugation

Detailed Description

The trial will compare the use of a microfluidic chip with the use of density gradient centrifugation for sperm preparation.

The hypothesis of the study is that the use of the microfluidic chip will improve the cumulative live birth rate of IVF compared to the density gradient method.

The trial will be conducted at the Centre of Assisted Reproduction and Embryology at Queen Mary Hospital and Kwong Wah Hospital. Infertile patients undergoing IVF treatment will be recruited during ovarian stimulation. They will be randomly assigned to one of two groups: the microfluidic chip group or the density gradient group. Both the patients and the clinicians will be blinded to the group allocation.

The inclusion criteria for the study are infertile women aged under 43 years undergoing ovarian stimulation for IVF. The exclusion criteria include women undergoing certain genetic testing, male factor infertility requiring surgical sperm retrieval, the use of donor oocytes and spermatozoa, certain uterine conditions, previous participation in the study, and participation in other randomized trials.

The eligible women will undergo standard IVF procedures, including ovarian stimulation, monitoring of follicle growth, trigger for oocyte retrieval, and oocyte retrieval itself. On the day of oocyte retrieval, the women will be randomly assigned to either the microfluidic chip group or the density gradient group for sperm preparation.

Semen samples will be collected on the day of oocyte retrieval and evaluated according to standard guidelines. Sperm DNA damage will also be assessed using an alkaline single-cell gel electrophoresis assay. Depending on the randomization, the sperm samples will be prepared either using the microfluidic chip or the density gradient centrifugation method.

After sperm preparation, oocytes will be fertilized using either conventional insemination or intracytoplasmic sperm injection. Fertilization will be confirmed by the presence of two pronuclei. Fresh embryo transfer will be performed 2-5 days after egg retrieval, and luteal phase support will be given. Excessive good quality embryos or blastocysts will be cryopreserved for future use.

For frozen embryo transfer, embryos or blastocysts will be replaced in subsequent natural or hormonal replacement cycles. Pregnancy will be confirmed by a urine pregnancy test, and transvaginal ultrasound will be performed to confirm viability and the number of fetuses.

Follow-up will be conducted to retrieve pregnancy and delivery data. The cumulative live birth rate will be calculated, and pregnancy outcomes, including birth weights and obstetric complications, will be recorded and compared between the two groups.

The study will continue until all cryopreserved embryos or blastocysts are used or until the participants become pregnant within 6 months after randomization. The pregnancy complications and congenital abnormalities will be monitored through hospital records or patient contact.

Ultimately, the study aims to determine whether the use of the microfluidic chip improves the cumulative live birth rate of IVF compared to the density gradient method.

• Study Type: Interventional
• Enrollment (Estimated): 1136
• Phase: Not Applicable

Contacts and Locations

• Study Contact: Name: YU WING TONG, MBBS; Phone Number: 92707722; Email: ptong@connect.hku.hk

Study Locations

• China
  • Hong Kong
    • Hong Kong, Hong Kong, China
      • Recruiting
      • Department of Obstetrics and Gynaecology
      • Contact: Ernest HY NG, MD; Phone Number: 852-22553400; Email: nghye@hku.hk
      • Principal Investigator: Ernest HY NG, MD

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Child; Adult
• Accepts Healthy Volunteers: No

Description

Inclusion Criteria:

• Infertile women aged <43 years at the time of ovarian stimulation for IVF

Exclusion Criteria:

• Women undergoing preimplantation genetic testing monogenic diseases, structural rearrangement of chromosomes or aneuploidy;
• Male factor requiring surgical sperm retrieval such as microscopic epididymal sperm aspiration and testicular sperm extraction;
• Use of donor oocytes and spermatozoa;
• Submucosal fibroid or hydrosalpinx shown on pelvic scanning and not surgically treated;
• Women who had been recruited into this study before and
• Women joining other randomized trials

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Treatment
• Allocation: Randomized
• Interventional Model: Parallel Assignment
• Masking: Quadruple

Number of Arms

2

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: microfluidic chip method; Microfluidic chip method has been used for sperm sorting in order to select the most motile and morphologically normal sperm for use in assisted reproductive technologies (ART) such as in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). In the microfluidic chip method for sperm sorting, a small amount of semen sample is loaded onto the chip, which contains channels and chambers that allow for the separation of sperm based on their motility and morphology. The chip is designed to mimic the natural environment of the female reproductive tract, where sperm undergo a series of selection processes before reaching the egg.	Device: Microfluidic chip; The Sperm Separation Device - ZyMōt Multi 850µL (ZyMōt Fertility, Inc) will be used. The microfluidics chamber will be used according to the manufacturer's instructions. 850 μL of the semen sample will be inserted into the inlet port of the device and 750 μL of fertilization media will be inserted into the outlet port. The device with the semen sample inside will be incubated in 6% CO2 at 37°C. After 30 min, 500 μL of the sample at the outlet port will be removed from the outlet port and pipetted into a labelled test tube.
Active Comparator: density-gradient centrifugation method; Density-gradient centrifugation is a commonly used method for sperm separation and purification. It is a technique that involves layering a semen sample on top of a gradient of different densities of a solution, typically a mixture of colloidal silica and sucrose, and then centrifuging the sample. The centrifugal force causes the sperm to migrate through the gradient, where they become separated based on their density.	Device: Density-gradient centrifugation; After liquefaction, sperm preparation will be completed by a discontinuous density gradient centrifugation method, using Pureception (CooperSurgical, Denmark) sperm density gradient media. The resulting sperm pellet after centrifugation will be washed once with the sperm washing medium (G-IVF Plus, Vitrolife, Sweden) The washed spermatozoa will be resuspended with the same medium, adjusting the final volume to 0.5 mL.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
cumulative live birth rate	cumulative live birth rate defined as the number of pregnancies leading to live birth within 6 months of randomisation.	within 6 months of randomisation.

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Live birth beyond 22 weeks of gestation per the first embryo transfer or FET	Live birth beyond 22 weeks of gestation per the first embryo transfer or FET	3 years
Positive urine pregnancy test per the first embryo transfer or FET	Positive urine pregnancy test per the first embryo transfer or FET	3 years
Clinical pregnancy per the first embryo transfer or FET defined as presence of intrauterine gestational sac on scanning at gestational week 6.	Clinical pregnancy per the first embryo transfer or FET defined as presence of intrauterine gestational sac on scanning at gestational week 6.	3 years
Ongoing pregnancy rate as presence of a fetal pole with pulsation at 8-10 weeks of gestation	Ongoing pregnancy rate as presence of a fetal pole with pulsation at 8-10 weeks of gestation	3 years
Miscarriage defined as a clinically recognized pregnancy loss before the 22 weeks of pregnancy and whose denominator is the clinical pregnancy.	Miscarriage defined as a clinically recognized pregnancy loss before the 22 weeks of pregnancy and whose denominator is the clinical pregnancy.	3 years
Multiple pregnancy: presence of more than one intrauterine sac at 6 weeks of gestation	Multiple pregnancy: presence of more than one intrauterine sac at 6 weeks of gestation	3 years
Ectopic pregnancy rate	Ectopic pregnancy rate	3 years

Collaborators and Investigators

• Sponsor: Professor Ernest Hung-Yu Ng

Study record dates

Study Major Dates

• Study Start (Actual): November 1, 2024
• Primary Completion (Estimated): November 30, 2027
• Study Completion (Estimated): November 30, 2029

Study Registration Dates

• First Submitted: June 29, 2023
• First Submitted That Met QC Criteria: August 16, 2023
• First Posted (Actual): August 22, 2023

Study Record Updates

• Last Update Posted (Estimated): December 6, 2024
• Last Update Submitted That Met QC Criteria: December 3, 2024
• Last Verified: December 1, 2024

More Information

Terms related to this study

• Keywords: microfluidic chip; density-gradient centrifugation; in-vitro fertilisation; sperm preparation
• Additional Relevant MeSH Terms: Urogenital Diseases; Genital Diseases; Infertility
• Other Study ID Numbers: UW 23-293

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: YES
• IPD Sharing Time Frame: after publication of the primary paper
• IPD Sharing Access Criteria: reasonable request
• IPD Sharing Supporting Information Type: STUDY_PROTOCOL; SAP; ICF; CSR

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: Yes
• product manufactured in and exported from the U.S.: Yes