Ice-free cryopreservation of whole pediatric testes for autologous banking and replantation.

整个儿科睾丸的无冰冷冻保存用于自体储存和再植。

• 批准号: 9919065
• 负责人: Kelvin G.M. Brockbank
• 金额: $ 30万
• 依托单位: TISSUE TESTING TECHNOLOGIES, LLC
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-15 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9919065
• 关键词: Achievement; Address; Adolescent; Adult; Age; AlamarBlue; Animals; Antioxidants; Aplastic Anemia; Apoptosis; Apoptotic; Applications Grants; Autoimmune Diseases; Autologous; Autologous Transplantation; Autopsy; Back; Benign; Biological Assay; Biopsy; Birth; Blood Vessel Tissue; Breeding; Cancer Patient; Cancer Survivor; Cell Survival; Cells; Characteristics; Child; Childhood; Chronic Childhood Arthritis; Clinic; Complex; Cooley' s anemia; Cryopreservation; Cryopreserved Cell; Cytotoxic Chemotherapy; Development; Diagnosis; Dimethyl Sulfoxide; Dissection; Dysmyelopoietic Syndromes; Equilibrium; Evaluation; Exposure to; Failure; Fanconi' s Anemia; Feasibility Studies; Female; Fertility; Fertilization; Fertilization in Vitro; Fibrosis; Formulation; Frequencies; Future; Genetic; Germ Cells; Germ Lines; Heart Valves; Heating; Hematological Disease; High Dose Chemotherapy; Histologic; Histology; Hormonal; Hormones; Human; Ice; Immune; In Vitro; Infertility; Injury; Interview; Klinefelter' s Syndrome; Lead; Letters; Libido; Licensing; Life; Macaca mulatta; Magnetism; Maintenance; Male Infertility; Malignant Neoplasms; Metals; Methods; Modeling; Nitrogen; Non-Malignant; Oocytes; Operative Surgical Procedures; Outcome; Parents; Patients; Phase; Pre-implantation Embryo Development; Pregnancy; Production; Propylene Glycols; Protocols documentation; Puberty; Public Health; Radiation therapy; Rattus; Reproductive Medicine; Rewarming; Rhesus; Sales; Sampling; Scrotum; Services; Sex Functioning; Sickle Cell Anemia; Skin; Small Business Innovation Research Grant; Spermatogenesis; Stains; Structure; Sucrose; Surgeon; Surgical Replantation; Systemic Lupus Erythematosus; Systemic Scleroderma; Technology; Testicular Tissue; Testing; Testis; Testosterone; Therapeutic immunosuppression; Time; Tissue Banks; Tissue Viability; Tissues; Transplantation; Trehalose; base; boys; cancer diagnosis; cancer therapy; chemotherapy; childhood cancer survivor; commercialization; cytopenia; cytotoxic; cytotoxicity; formamide; implantation; improved; innovation; male; nanoparticle; nanowarming; novel; prepuberty; preservation; radio frequency; repaired; reproductive; reproductive organ; restoration; sperm cell; stem cells; vapor; ward; young adult

Thousands of young males under the age of 20 are diagnosed with cancer each year. The overall cure rates of these cancer patients are approaching 80%; therefore, the number of childhood cancer survivors is increasing over time. It is well documented that cancer treatments may result in temporary, long-term, or permanent gonadal failure in male cancer survivors. Certain benign hematological disorders, such as myelodysplasia, sickle cell disease, aplastic anemia, thalassemia major, and Fanconi anemia, and severe autoimmune diseases unresponsive to immunosuppressive therapy, such as juvenile idiopathic arthritis, juvenile systemic lupus erythematosus, systemic sclerosis and immune cytopenias, necessitate administration of high dose chemotherapy that may also impact testicles. In immature boys, in particular, storing testicular tissue prior to such cytotoxic treatments for future replantation could be an option. This proposal specifically addresses cryopreservation of pediatric testes. This male reproductive organ is contained within the scrotum, and is responsible for the production of sperm and the male hormone, testosterone that is pivotal for the maintenance of male sexual characteristics. As such, injury to the testes can render a young male infertile and needing administration of exogenous testosterone. Klinefelter’s syndrome patients would also benefit. Klinefelter’s syndrome the most common (1:500 new male born) genetic reason for male infertility, suffer from progressive germ cell loss and testis fibrosis with onset of puberty. We propose to remedy this by cryopreservation of testes that can be used for restoration of sexual function, fertility and hormonal balance in males before they are damaged. A single testis from each patient would be cryopreserved. This would permit either replantation of the patient’s intact testis, implantation of viable spermatogonial stem cells in the remaining testis, or ectopic implantation of functional spermatogonial stem cells. Each strategy would be capable of completing spermatogenesis and provide testosterone in a regulated way. We propose a novel innovative approach using ice-free vitrification to cryopreserve pediatric testes employing a young rat model. No animals will be sacrificed because we are using heterozygous and wild type culls from breeding colonies post-mortem. In this Phase I SBIR proposal in vitro methods and histology will be used to assess outcomes in three specific aims. Feasibility for progression to a Phase II SBIR grant proposal will be demonstrated by achievement of >80% cell viability by 3 methods and <20% apoptotic cells after 3 months of storage in vapor phase nitrogen. We have already assembled a world-class team of experts in surgery and reproductive medicine to guide the Company in Phase II and Phase III commercialization efforts.

每年，成千上万的20岁以下男性被诊断出患有癌症。这些癌症患者的总体治愈率接近80％。因此，随着时间的流逝，儿童癌症存活的数量正在增加。有充分的文献证明，癌症治疗可能导致男性癌症存活中的暂时，长期或永久性性腺衰竭。某些良性血液学疾病，例如骨髓增生，镰状细胞疾病，性障碍性贫血，thalassyapia joart和fanconi贫血以及严重的自身免疫性疾病对免疫抑制治疗无反应症免疫细胞减少症，必要的高剂量化学疗法，这也可能影响睾丸。在未成熟的男孩中，尤其是在这种细胞毒性治疗之前存储测定的组织以进行未来再生。提案专门涉及小儿测试的冷冻保存。这种雄性生殖器官包含在阴囊内，负责精子和雄性马龙的产生，这是维持男性性特征的关键的睾丸激素。因此，对睾丸的损伤会导致年轻的男性不育，需要给予外源睾丸激素。 KlineFelter综合征患者也将受益。 Klinefelter综合征是男性不育症的最常见（1：500新的男性出生）遗传学原因，患有渐进的生殖细胞损失和睾丸纤维化，青春期发作。我们建议通过冷冻保存测试来记住这一点，这些测试可用于恢复男性的性功能，生育能力和痛苦。来自每个患者的单个睾丸将是冷冻保存的。这将允许重新补充患者的完整睾丸，在其余睾丸中植入可行的精子干细胞，或者植入功能性精子干细胞的生态植入。每种策略将能够完成精子发生并以调节的方式提供睾丸激素。我们提出了一种使用无冰玻璃化的新型创新方法，用于使用年轻大鼠模型的冷冻保存小儿测试。由于我们在验尸后使用杂合和野生型杯子，因此不会牺牲任何动物。在这一阶段，I SBIR建议的体外方法和组织学将用于评估三个特定目标的结果。通过3种方法实现> 80％的细胞活力，在蒸气期氮的储存3个月后，将证明> 80％的细胞活力，<20％的凋亡细胞实现> 80％的细胞活力，将证明其前进的可行性。我们已经组建了一个世界一流的手术和生殖医学专家团队，以指导公司在第二阶段和第三阶段的商业化工作中。