Testicular Tissue Freezing for Infertility

(TTC Trial)

Overseen By Leslie Appiah, MD

Age: Any Age

Sex: Male

Trial Phase: Academic

Sponsor: University of Colorado, Denver

Disqualifiers: Psychological conditions, Anesthesia risk, others

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

Approved in 2 Jurisdictions This treatment is already approved in other countries

Compensation: Varies

Number of Visits: 1

60 Participants Needed

Trial Summary

What is the purpose of this trial?

The "Testicular Tissue Cryopreservation" study is open to a subset of patients facing disease or treatment regimens that could lead to infertility (gonadotoxic therapies). For some of these patients, experimental testicular tissue cryopreservation is the only fertility preservation option available. The overall objective of this study is to determine the feasibility and acceptability of testicular tissue cryopreservation in male patients of all ages who have a condition or will undergo a treatment that can cause infertility.

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 Testicular Tissue Cryopreservation for infertility?

Research shows that testicular tissue cryopreservation can preserve fertility in prepubertal boys who cannot store sperm due to cancer treatments or genetic conditions. While successful in animals, human applications are still experimental, but advancements in lab techniques are improving the chances of producing fertile sperm from preserved tissue. ^{1 2 3 4 5}

Is testicular tissue freezing safe for humans?

Cryopreservation of testicular tissue is considered a low-risk procedure, especially for pre-pubertal and adolescent boys at risk of infertility due to cancer treatments, with minimal complications reported. ^{1 6 7 8 9}

How is Testicular Tissue Cryopreservation different from other treatments for infertility?

Testicular Tissue Cryopreservation is unique because it involves freezing testicular tissue to preserve fertility, especially for pre-pubertal boys who cannot produce sperm yet. Unlike other treatments, it focuses on preserving spermatogenic cells (cells that can develop into sperm) before they are damaged by treatments like chemotherapy. ^{4 10 11 12 13}

Research Team

Leslie Appiah, MD

Principal Investigator

University of Colorado, Denver

Eligibility Criteria

This trial is for male patients of any age who may become infertile due to a medical condition or treatments that are harmful to reproductive cells. It's specifically aimed at those for whom freezing testicular tissue is the only option to preserve fertility.

Inclusion Criteria

I am a male.

In significant risk of infertility as defined by: Cyclophosphamide equivalent dose (CED) ≥4 g/m2, Total body irradiation (TBI), Testicular radiation >2.5 Gy, Cisplatin 500 mg/m2, Bone Marrow Transplant (BMT), Or have a medical condition or malignancy that requires removal of all or part of one or both testicles, Or have newly diagnosed or recurrent disease. Those who were not enrolled at the time of initial diagnosis (i.e., patients with recurrent disease) are eligible if they have not previously received therapy that is viewed as likely to result in complete and permanent loss of testicular function, Have two testicles if undergoing elective removal of a testicle for fertility preservation only. Note: removal of both testicles will limit fertility preservation options, Sign an approved informed consent and authorization permitting the release of personal health information. The patient and/or the patient's legally authorized guardian must acknowledge in writing that consent for specimen collection has been obtained, in accordance with institutional policies approved by the U.S. Department of Health and Human Services, Consent for serum tests for infectious diseases [including Hepatitis B Surface Antigen, HCV Antibody, and Human Immunodeficiency Virus (HIV) AG/AB Screen] to be performed at the time of testicular tissue harvesting, Undergo a full history and physical examination and obtain standard pre-operative clearance (based on the most recent ACC/AHA Guideline for Perioperative Cardiovascular Evaluation for Noncardiac Surgery) as determined by their primary surgeon

I am scheduled for treatment that may permanently affect my ability to produce sperm.

Exclusion Criteria

I have a health condition that makes anesthesia and surgery riskier for me.

Diagnosed with psychological, psychiatric, or other conditions which prevent giving fully informed consent

Timeline

Screening

Participants are screened for eligibility to participate in the trial

2-4 weeks

Testicular Tissue Cryopreservation

Participants undergo a testicular tissue biopsy for cryopreservation

Up to 48 hours

1 visit (in-person)

Post-procedure Monitoring

Participants are monitored for safety, including adverse events such as bleeding

24 hours

Follow-up

Participants are monitored for long-term safety and effectiveness, including assessment of long-term pain

6 months

Long-term Acceptability Assessment

Participants' acceptability of the procedure is assessed annually using the Decision Regret Scale

Annually until the age of majority

Treatment Details

Interventions

• Testicular Tissue Cryopreservation

Trial Overview The study focuses on taking a small sample of testicular tissue and freezing it (cryopreservation) with the goal of preserving future fertility. The feasibility and acceptability of this procedure in patients at risk of infertility will be evaluated.

Participant Groups

Group I: Testicular Tissue Cryopreservation Experimental Treatment 1 Intervention

Enrolled participants will undergo a testicular tissue biopsy. The method and amount of tissue procurement will be at the discretion of the surgeon.

Testicular Tissue Cryopreservation is already approved in United States, European Union for the following indications:

Approved in United States as Testicular Tissue Cryopreservation for:

• Experimental fertility preservation for prepubertal males facing gonadotoxic treatments or diseases

Approved in European Union as Testicular Tissue Cryopreservation for:

• Experimental fertility preservation for prepubertal males facing gonadotoxic treatments or diseases

Who Is Running the Clinical Trial?

University of Colorado, Denver

Lead Sponsor

Trials

1,842

Recruited

3,028,000+

Findings from Research

Uncontrolled slow freezing (USF) using 1.5 M DMSO and 0.15 M sucrose is an effective and user-friendly method for cryopreserving human testicular tissue, showing better preservation of cell structure and function compared to conventional controlled slow freezing (CSF) protocols.

In a study involving 160 testicular tissue fragments from 14 patients, USF significantly reduced cell death and maintained the integrity of the seminiferous epithelium and spermatogonial cells, making it a promising alternative for fertility preservation, especially in resource-limited settings.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

What is the best cryopreservation protocol for human testicular tissue banking? Baert, Y., Van Saen, D., Haentjens, P., et al.[2022]

Cryopreservation of testicular cells and tissues is a promising method for preserving fertility in pre-pubertal males undergoing treatments that may harm their ability to produce sperm, with advancements in in vitro spermatogenesis enhancing the potential for producing viable sperm from these preserved tissues.

Recent developments in testicular tissue engineering and in vitro sperm production from cryopreserved stem cells have reduced the need for transplantation, offering new hope for fertility restoration in males affected by gonadotoxic treatments.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Strategies for cryopreservation of testicular cells and tissues in cancer and genetic diseases. Patra, T., Pathak, D., Gupta, MK.[2021]

Testicular tissue cryopreservation is a promising fertility preservation method for prepubertal males, but there is currently no standardized procedure for effectively freezing immature testicular tissue.

A review of 11 studies revealed that different cryopreservation protocols vary in freezing rates and cryoprotectant media, indicating that further research is needed to identify the best methods for preserving tissue integrity and restoring fertility.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Freezing protocols for the cryopreservation of immature testicular tissue - a systematic review. [2022]

A study involving 34 boys at risk of fertility loss due to cancer treatment showed that most had normal or increased germ cell counts in their testicular tissue, indicating potential for fertility preservation through cryopreservation.

However, abnormal maturation of germ cells was observed in some older patients, suggesting that further research is needed to understand the implications for fertility preservation in this population.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Histology of Testicular Biopsies Obtained for Experimental Fertility Preservation Protocol in Boys with Cancer. Pietzak, EJ., Tasian, GE., Tasian, SK., et al.[2018]

Testicular tissue cryopreservation is a promising method for preserving spermatogonial stem cells (SSCs) in pre-pubertal boys and adolescents at risk of infertility due to cancer treatment or genetic disorders, as traditional sperm cryopreservation is not viable for these groups.

Establishing a testicular tissue banking system is crucial for advancing SSC transplantation in humans, which has shown success in other species but remains experimental in people, highlighting the need for a multidisciplinary approach to ensure safe and effective implementation.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Experimental testicular tissue banking to generate spermatogenesis in the future: A multidisciplinary team approach. Sadri-Ardekani, H., McLean, TW., Kogan, S., et al.[2022]

Cryopreservation of testicular tissue (TT) in pre-pubertal boys is a safe procedure, with only 5.9% of patients experiencing complications, such as resolved epididymo-orchitis and a conservatively managed torsed appendix testis.

The study involved 34 patients with an average age of 6.9 years, and most biopsies were performed alongside other necessary medical procedures, indicating that TT biopsy can be integrated into the care of young patients at risk of infertility due to cancer treatment.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Cryopreservation of testicular tissue in pre-pubertal and adolescent boys at risk for infertility: A low risk procedure. Ming, JM., Chua, ME., Lopes, RI., et al.[2019]

Orthotopic grafting of frozen-thawed human testicular tissue to mouse testes resulted in high survival rates of spermatogonial stem cells, indicating a promising method for fertility preservation in patients undergoing gonadotoxic treatments.

Grafts from older boys (ages 12 and 13) showed not only higher germ cell survival but also differentiation into primary and secondary spermatocytes, while grafts from younger boys (ages 3 and 5) did not differentiate, suggesting that donor age significantly impacts the success of testicular tissue grafting.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Meiotic activity in orthotopic xenografts derived from human postpubertal testicular tissue. Van Saen, D., Goossens, E., Bourgain, C., et al.[2011]

Cryopreservation of sperm is the standard method for preserving fertility in men before undergoing treatments that may impair fertility, with various techniques available for sperm collection depending on individual circumstances, such as azoospermia or retrograde ejaculation.

In Germany, strict regulations govern the cryopreservation process, requiring specific approvals for both sperm and testicular tissue preservation, ensuring safety and compliance in fertility preservation practices.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

[Fertility and fertility preservation in men]. Hoffmann, I., Greither, T., Behre, HM.[2023]

Testicular tissue cryopreservation (TTC) was performed safely in 23 male patients aged 5 months to 18 years, with a majority showing normal testicular tissue and germ cells after biopsy, indicating potential for future fertility preservation.

The procedure did not delay the start of necessary medical treatments, and only one case of postoperative infection was reported, suggesting that TTC can be integrated effectively into the treatment plans for children at risk of azoospermia.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Testicular wedge biopsy for fertility preservation in children at significant risk for azoospermia after gonadotoxic therapy. Corkum, KS., Lautz, TB., Johnson, EK., et al.[2019]

In a study involving testicular tissue from 16 infertile men, DMSO was found to be the most effective cryoprotectant, preserving 70% of the seminiferous tubules' structure after freezing, compared to only 37% with 1,2-propanediol and severe damage with glycerol.

The research indicates that DMSO not only maintains the structural integrity of spermatogonia but also preserves the function of Leydig cells, which continue to release testosterone after cryopreservation.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Optimizing cryopreservation of human testicular tissue: comparison of protocols with glycerol, propanediol and dimethylsulphoxide as cryoprotectants. Keros, V., Rosenlund, B., Hultenby, K., et al.[2013]

Systematic cryo-banking of testicular tissues can significantly enhance the management of male fertility and help preserve rare mammalian genotypes by allowing for the development of viable sperm from preserved germ cells.

Current methods for obtaining and preserving testicular tissues include biopsies from living individuals and various cryopreservation techniques, but the process of obtaining functional sperm still requires optimization, particularly through xenografting into immunodeficient mice.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Cryopreservation and Culture of Testicular Tissues: An Essential Tool for Biodiversity Preservation. Silva, AMD., Pereira, AF., Comizzoli, P., et al.[2021]

In a study involving 87 (pre)pubertal boys with cancer, a control slow freezing protocol was found to effectively preserve the architecture and integrity of testicular tissue after thawing, with minimal cryodamage observed.

The number of spermatogonia and the expression of DNA repair markers remained stable post-thawing, indicating that the freezing process does not significantly impact the potential for future fertility preservation.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Assessment of the architecture and integrity of frozen-thawed testicular tissue from (pre)pubertal boys with cancer. Rives-Feraille, A., Liard, A., Bubenheim, M., et al.[2022]

Testicular tissue from pre-pubertal boys can be successfully cryopreserved using a slow programmed freezing method with 5% dimethyl sulphoxide (DMSO) as a cryoprotectant, preserving the integrity of key cell types necessary for future fertility.

The study found that while the freezing protocol developed for adult testicular tissue was effective, a protocol designed for fetal tissue caused more damage, highlighting the importance of optimizing cryopreservation techniques specifically for pre-pubertal testicular tissue.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Methods of cryopreservation of testicular tissue with viable spermatogonia in pre-pubertal boys undergoing gonadotoxic cancer treatment. Keros, V., Hultenby, K., Borgström, B., et al.[2022]

References

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

What is the best cryopreservation protocol for human testicular tissue banking? [2022]

2. Germany pubmed.ncbi.nlm.nih.gov

Strategies for cryopreservation of testicular cells and tissues in cancer and genetic diseases. [2021]

3. England pubmed.ncbi.nlm.nih.gov

Freezing protocols for the cryopreservation of immature testicular tissue - a systematic review. [2022]

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

Histology of Testicular Biopsies Obtained for Experimental Fertility Preservation Protocol in Boys with Cancer. [2018]

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

Experimental testicular tissue banking to generate spermatogenesis in the future: A multidisciplinary team approach. [2022]

6. England pubmed.ncbi.nlm.nih.gov

Cryopreservation of testicular tissue in pre-pubertal and adolescent boys at risk for infertility: A low risk procedure. [2019]

7. England pubmed.ncbi.nlm.nih.gov

Meiotic activity in orthotopic xenografts derived from human postpubertal testicular tissue. [2011]

8. Germany pubmed.ncbi.nlm.nih.gov

[Fertility and fertility preservation in men]. [2023]

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

Testicular wedge biopsy for fertility preservation in children at significant risk for azoospermia after gonadotoxic therapy. [2019]

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

Optimizing cryopreservation of human testicular tissue: comparison of protocols with glycerol, propanediol and dimethylsulphoxide as cryoprotectants. [2013]

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

Cryopreservation and Culture of Testicular Tissues: An Essential Tool for Biodiversity Preservation. [2021]

12. England pubmed.ncbi.nlm.nih.gov

Assessment of the architecture and integrity of frozen-thawed testicular tissue from (pre)pubertal boys with cancer. [2022]

13. England pubmed.ncbi.nlm.nih.gov

Methods of cryopreservation of testicular tissue with viable spermatogonia in pre-pubertal boys undergoing gonadotoxic cancer treatment. [2022]