Vitrification Systems for Egg Freezing

Overseen By M Ángeles Romero, Bachelors Degree on Bioligy

Age: 18 - 65

Sex: Female

Trial Phase: Phase 2 & 3

Sponsor: Overture Life

Disqualifiers: Age, Bmi, Uterine pathologies, others

No Placebo Group All trial participants will receive the active study treatment (no placebo)

Prior Safety Data This treatment has passed at least one previous human trial

Compensation: Varies

Number of Visits: Unknown

260 Participants Needed

Trial Summary

What is the purpose of this trial?

This study will recruit 260 female recipients (and up to a maximum of 130 donors). This study is a sequel to the protocol CP-DV-000-GEN-003, entitled, "NON-INFERIORITY STUDY COMPARING OVERTURE SEMI-AUTOMATED VITRIFICATION SYSTEM ("DaVitri") TO STANDARD MANUAL PROCESS USING BLASTOCYST RATE AS THE PRIMARY END POINT". Donors will provide informed consent and will be enrolled before egg retrieval. Once the oocytes are retrieved, they will undergo thorough denudation to remove corona cells and will be assessed for maturity. Subsequently, mature MII oocytes exhibiting good morphology, as per the Vienna Consensus, ranging between 12 and 30 in number, will be processed. These oocytes will then be randomly divided into groups comprising 6 to 8 oocytes each. Therefore, from a single donor, 2 to 4 groups can be generated. An identifier will be assigned for each group. One of the groups will be randomly selected. This group will be randomly assigned to DaVitri or Control. The remaining groups will be randomly selected and assigned alternatively to Control and DaVitri groups: Control oocytes will be manually vitrified and placed into cryogenic storage. Test oocytes will be prepared for vitrification using the DaVitri system. Following preparation with DaVitri, test oocytes will be loaded onto a cryogenic device that is placed into cryogenic storage. The same preservation media (Kitazato) will be used to prepare all oocytes. All oocytes will be warmed manually. Survival rate will be recorded following warming. After warming, the oocytes will be fertilized via Intracytoplasmic Sperm Injection (ICSI) with either donor or patient's partner sperm and resulting embryos cultured to blastocyst stage, keeping score of which embryos come from the Test or Control group. Fertilization rate will be recorded. Embryos will be morphologically assessed (according to Gardner grading system) on day 5-6 to determine blastulation rates and embryo quality in both groups. All the embryos will be vitrified according to the clinic\'s routine process and stored for further recipients. Recipients will provide informed consent and will be enrolled before the retrieval of donor eggs. Once the donor has been matched with the recipient according to the phenotypic, demographic characteristics (following the regular clinical process established in the clinic donation program), the group of oocytes assigned will be randomly selected from either the DaVitri processed group or the manually processed group. Recipients of donated eggs will receive a single embryo transfer (SET). The primary endpoint is Clinical Pregnancy Rate. Clinical Pregnancy will be confirmed by the presence of sac in uterus and chemical confirmation at 6-7 weeks after embryo transference, via ultrasound. Only the first embryo transfers will be used to calculate the primary endpoint. Successive transfers of any embryo group will not be considered inside the study.

Will I have to stop taking my current medications?

The trial information does not specify whether you need to stop taking your current medications. It's best to discuss this with the trial coordinators or your doctor.

What data supports the effectiveness of the treatment Oocyte Cryopreservation for egg freezing?

Research shows that improvements in vitrification (a fast freezing method) have significantly increased pregnancy rates over the past five years. Closed vitrification systems, which avoid direct contact with liquid nitrogen, have been validated for routine use in egg donation programs, with higher oocyte survival rates linked to increased chances of pregnancy. ^{1 2 3 4 5}

Is egg freezing (oocyte vitrification) safe for humans?

Oocyte vitrification, a method used for egg freezing, appears to be safe for both the eggs and the resulting children in the short term, according to studies involving over 4,000 babies. However, more long-term data is needed to fully understand its safety. ^{6 7 8 9 10}

How does the treatment Oocyte Cryopreservation differ from other treatments for egg freezing?

Oocyte Cryopreservation, or egg freezing, is unique because it uses a process called vitrification, which involves ultra-rapid cooling to prevent ice crystal formation that can damage the eggs. This method is distinct from traditional slow-freezing techniques and is designed to preserve the quality and viability of the eggs more effectively. ^{11 12 13 14 15}

Eligibility Criteria

This trial is for up to 260 women receiving egg donations and a maximum of 130 donors. Donors must provide mature, good quality eggs, while recipients are matched based on phenotypic and demographic characteristics. Participants need to consent to the use of their eggs or embryos in the study.

Inclusion Criteria

I am a healthy female aged 18-35 with an AMH level between 2-10 ng/ml.

I am a woman aged 18-45, using a donated egg for IVF, with a normal uterus, planning to transfer one embryo.

Exclusion Criteria

Age >35 years of age BMI <18.5 or >25 Infertility history Three previously failed IVF cycles, including abortions Abnormal ovulation cycle FSH >10 or AMH <2 Alcoholism, drug addiction Infectious diseases (such as HIV, hepatitis, sexually transmitted infections) Donors producing less than 12 mature oocytes of good quality (according to Vienna Consensus criteria) (donors)

Severe male factor Infertility (unless donated sperm is used) Alcoholism, drug addiction (sperm)

Age >45 years of age BMI <18.5 or >29 Patients using a surrogate Evidence of uterine pathologies Severe male factor Infertility (unless donated sperm is used) Alcoholism, drug addiction Infectious diseases (such as HIV, hepatitis, sexually transmitted infections) (recipients)

Timeline

Screening

Participants are screened for eligibility to participate in the trial

2-4 weeks

Oocyte Retrieval and Vitrification

Donors provide oocytes which are then vitrified using either the DaVitri system or manually, and stored in cryogenic storage.

2 weeks

Fertilization and Embryo Culture

Oocytes are warmed, fertilized via ICSI, and cultured to the blastocyst stage. Embryos are assessed for quality and vitrified for future transfer.

2 weeks

Embryo Transfer and Initial Pregnancy Assessment

Recipients receive a single embryo transfer, and clinical pregnancy is confirmed by ultrasound and chemical tests at 6-7 weeks post-transfer.

6-7 weeks

Follow-up

Participants are monitored for ongoing pregnancy and delivery success, with assessments for congenital abnormalities and delivery outcomes.

9 months

Treatment Details

Interventions

• Oocyte Cryopreservation

Trial Overview The study compares two methods of freezing eggs: DaVitri (a semi-automated system) versus manual vitrification. Eggs from each donor are split into groups and randomly assigned to either method before being fertilized and grown into embryos for transfer.

Participant Groups

Group I: Test Oocytes Experimental Treatment 1 Intervention

Test oocytes will be prepared for vitrification using the DaVitri system. Following preparation with DaVitri, test oocytes will be loaded onto a cryogenic device that is placed into cryogenic storage.

Group II: Manually vitrified oocytes Active Control 1 Intervention

Control oocytes will be manually vitrified and placed into cryogenic storage.

Who Is Running the Clinical Trial?

Overture Life

Lead Sponsor

Trials

5

Recruited

960+

Findings from Research

Oocyte cryopreservation using vitrification is significantly more effective than slow freezing, showing higher survival, implantation, and pregnancy rates based on data from the Italian National Assisted Reproductive Technology Register covering 2009-2014.

Despite the advantages of vitrification, the success rates can vary widely among different centers, and transferring fresh or cryopreserved embryos still yields better outcomes than using embryos from cryopreserved oocytes, except in highly experienced centers.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Evolution of human oocyte cryopreservation: slow freezing versus vitrification. Levi-Setti, PE., Patrizio, P., Scaravelli, G.[2018]

The study found that using closed vitrification systems for egg donation significantly predicts oocyte survival and clinical pregnancy rates, with each additional vitrified oocyte increasing survival odds by 15%.

The oocyte survival rate directly correlates with clinical pregnancy probability, showing that a 100% survival rate leads to a 32% chance of clinical pregnancy, supporting the routine use of closed vitrification systems in egg donation programs.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Closed vitrification system and egg donation: Predictive factors of oocyte survival and pregnancy. Gala, A., Ferrières-Hoa, A., Loup-Cabaniols, V., et al.[2021]

Oocyte cryopreservation (OC) has significantly improved in effectiveness over the past five years due to advancements in vitrification technology, leading to higher pregnancy rates for women using this method.

OC is a valuable option for women facing fertility threats from medical treatments or age-related decline, and it can also enhance donor egg programs and provide alternatives for couples concerned about ethical issues with embryo preservation.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Oocyte cryopreservation: advances and drawbacks. Dovey, S.[2012]

A study involving 1,027 children from vitrified oocytes and 1,224 from fresh oocytes found that oocyte vitrification does not negatively impact obstetric and perinatal outcomes, such as birth weight and rates of complications.

While there were more invasive procedures and fewer urinary tract infections in the vitrified oocyte group, these findings suggest that vitrification is a safe option for assisted reproduction without increasing risks for mothers or babies.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Obstetric and perinatal outcome of babies born from vitrified oocytes. Cobo, A., Serra, V., Garrido, N., et al.[2022]

A large clinical trial involving 600 recipients found that vitrified (frozen) oocytes have a comparable ongoing pregnancy rate (43.7%) to fresh oocytes (41.7%), demonstrating the effectiveness of oocyte cryopreservation in assisted reproductive technology (ART).

The study confirmed the non-inferiority of vitrified oocytes compared to fresh ones, suggesting that using frozen eggs can be a viable alternative without compromising pregnancy outcomes, which could expand options for egg banking programs.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Use of cryo-banked oocytes in an ovum donation programme: a prospective, randomized, controlled, clinical trial. Cobo, A., Meseguer, M., Remohí, J., et al.[2022]

A systematic review of 13 observational studies involving 4,159 babies suggests that oocyte vitrification is a safe method for cryopreservation, with no significant negative impacts on neonatal health outcomes in the short term.

Despite the positive findings, the review highlights the need for more long-term studies and randomized controlled trials to better understand the potential long-term effects of oocyte vitrification on child health.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

The Impact of Oocyte Vitrification on Offspring: a Systematic Review. Da Luz, CM., Caetano, MA., Berteli, TS., et al.[2022]

Fertility preservation through ovarian stimulation and cryopreservation is generally safe and can be completed in about 2 weeks, without negatively impacting disease severity or long-term survival for patients with serious medical conditions.

Modern techniques allow for high oocyte yield and can be initiated at various points in the menstrual cycle, with over 25% of patients returning to use fertility services later, highlighting the effectiveness of these preservation methods.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Oocyte and embryo cryopreservation before gonadotoxic treatments: Principles of safe ovarian stimulation, a systematic review. Ozcan, MC., Snegovskikh, V., Adamson, GD.[2022]

Vitrification of mature oocytes and embryos leads to better clinical outcomes in female fertility preservation without increasing risks of DNA damage or other genetic issues compared to traditional freezing methods.

The review analyzed data from studies published between 1980 and 2013, focusing on various cryopreservation techniques, and concluded that both embryo and oocyte vitrification are safe and effective options for preserving female fertility.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Female fertility: is it safe to "freeze?". Zhang, L., Yan, LY., Zhi, X., et al.[2019]

The review provides evidence-based guidelines on the effectiveness and safety of human oocyte cryopreservation methods, specifically slow freezing and vitrification, focusing on factors like oocyte maturity and cryopreservation techniques.

Key outcome measures such as cryosurvival, fertilization rates, and clinical pregnancy rates were examined, highlighting the importance of these factors in determining the success of oocyte freezing and subsequent pregnancies.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Human oocyte cryopreservation: evidence for practice. Cutting, R., Barlow, S., Anderson, R.[2009]

A survey conducted by the UK Association of Clinical Embryologists revealed a higher incidence of adverse events related to cryopreserved material in IVF units than previously thought, indicating a significant risk to both personnel and stored samples.

The study emphasizes the urgent need for improved training and awareness among staff, as well as a coordinated effort to collect and share data on adverse events to enhance safety and risk assessment in cryostorage practices.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Risks associated with cryopreservation: a survey of assisted conception units in the UK and Ireland. Tomlinson, M., Morroll, D.[2016]

Vitrifying ovarian tissue using only ethylene glycol (EG) as a cryoprotectant preserves the structure of ovarian follicles as effectively as using a combination of multiple cryoprotectants, based on analysis of tissue from women undergoing cesarean sections.

No signs of apoptosis were found in primordial and primary follicles after vitrification and warming, indicating that both methods are safe for preserving ovarian tissue for potential future use.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Preservation of human ovarian follicles within tissue frozen by vitrification in a xeno-free closed system using only ethylene glycol as a permeating cryoprotectant. Sheikhi, M., Hultenby, K., Niklasson, B., et al.[2013]

The novel automatic vitrification device demonstrated high survival rates for cryopreserved oocytes and embryos, with 95% survival for mice MII oocytes and 97% for blastocysts after rewarming.

This device could standardize and simplify the vitrification process, making it essential for the increasing global demand for cryopreservation of reproductive tissues.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

A new, simple, automatic vitrification device: preliminary results with murine and bovine oocytes and embryos. Arav, A., Natan, Y., Kalo, D., et al.[2019]

The new clean liquid air (CLAir) system and sterile storage canister (Esther) demonstrated similar cooling rates to liquid nitrogen while significantly reducing contamination risks, as shown by bioburden tests that revealed no contamination in CLAir and Esther compared to high contamination in commercial liquid nitrogen.

Vitrification of both mouse embryos and human oocytes using CLAir resulted in over 90% survival rates and 100% survival after three cycles, indicating that this method is as effective as traditional liquid nitrogen while being safer.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

New methods for cooling and storing oocytes and embryos in a clean environment of -196°C. Arav, A., Natan, Y., Levi-Setti, PE., et al.[2017]

Vitrification has become the standard method for cryopreserving bovine oocytes due to their high susceptibility to chilling injury, but a consistent method for achieving high blastocyst development rates is still lacking, with rates often exceeding 10% but showing high variability.

Porcine oocytes are particularly challenging to cryopreserve, with few studies achieving more than 5% blastocyst development; however, applying hydrostatic pressure before vitrification has shown promise in improving outcomes, with some treatments resulting in over 10% blastocyst development.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

A chronologic review of mature oocyte vitrification research in cattle, pigs, and sheep. Mullen, SF., Fahy, GM.[2012]

Cryopreservation of human oocytes has improved significantly, providing a valuable option for preserving fertility in individuals facing risks like cancer treatments or advanced maternal age, but challenges such as cryodamage still exist.

The review highlights the need for further research into optimizing vitrification and warming techniques, including the selection of cryoprotectants and cooling rates, to enhance the success rates of oocyte cryopreservation.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Theoretical and experimental basis of oocyte vitrification. Smith, GD., Motta, EE., Serafini, P.[2011]

References

1. England pubmed.ncbi.nlm.nih.gov

Evolution of human oocyte cryopreservation: slow freezing versus vitrification. [2018]

2. France pubmed.ncbi.nlm.nih.gov

Closed vitrification system and egg donation: Predictive factors of oocyte survival and pregnancy. [2021]

3. Italy pubmed.ncbi.nlm.nih.gov

Oocyte cryopreservation: advances and drawbacks. [2012]

4. United States pubmed.ncbi.nlm.nih.gov

Obstetric and perinatal outcome of babies born from vitrified oocytes. [2022]

5. England pubmed.ncbi.nlm.nih.gov

Use of cryo-banked oocytes in an ovum donation programme: a prospective, randomized, controlled, clinical trial. [2022]

6. United States pubmed.ncbi.nlm.nih.gov

The Impact of Oocyte Vitrification on Offspring: a Systematic Review. [2022]

7. United States pubmed.ncbi.nlm.nih.gov

Oocyte and embryo cryopreservation before gonadotoxic treatments: Principles of safe ovarian stimulation, a systematic review. [2022]

8. China pubmed.ncbi.nlm.nih.gov

Female fertility: is it safe to "freeze?". [2019]

9. England pubmed.ncbi.nlm.nih.gov

Human oocyte cryopreservation: evidence for practice. [2009]

10. England pubmed.ncbi.nlm.nih.gov

Risks associated with cryopreservation: a survey of assisted conception units in the UK and Ireland. [2016]

11. United States pubmed.ncbi.nlm.nih.gov

Preservation of human ovarian follicles within tissue frozen by vitrification in a xeno-free closed system using only ethylene glycol as a permeating cryoprotectant. [2013]

12. Netherlands pubmed.ncbi.nlm.nih.gov

A new, simple, automatic vitrification device: preliminary results with murine and bovine oocytes and embryos. [2019]

13. Netherlands pubmed.ncbi.nlm.nih.gov

New methods for cooling and storing oocytes and embryos in a clean environment of -196°C. [2017]

14. United States pubmed.ncbi.nlm.nih.gov

A chronologic review of mature oocyte vitrification research in cattle, pigs, and sheep. [2012]

15. Netherlands pubmed.ncbi.nlm.nih.gov

Theoretical and experimental basis of oocyte vitrification. [2011]