Gene Abnormality, Infertility, Male, Sterility, Male
Conditions
Brief summary
Purpose: This clinical trial aims to explore the potential for human sperm production in vitro by sustaining a laboratory-cultured adult testicular environment. It also seeks to identify genetic factors contributing to human sterility and failed spermatogenesis. The study's primary objectives include: 1. Identifying genomic markers associated with sterility and failed spermatogenesis. 2. Developing an ex vivo (outside the body) testis organ-on-a-chip (iTestis) to support stem cell cultivation. 3. Determining whether human spermatogenesis can be re-created in vitro using stem cells nurtured in the iTestis model. Study Description: Researchers will analyze the genomic profiles of fertile and sterile male participants to map genetic abnormalities associated with sterility. Using testicular and skin tissue samples from participants, spermatogonial stem cells and pluripotent stem cells will be isolated and utilized to construct the ex vivo iTestis. This system will integrate genomic insights and prior research to foster human spermatogenesis outside the body. Participant Involvement: Participants will provide the following samples: * Blood sample for serum analysis. * A skin tissue biopsy. * Testicular tissue, obtained through fine needle aspiration (FNA) or testicular sperm extraction (TESE), as part of a routine procedure. All procedures will be conducted by the principal investigator and qualified research staff, ensuring participant safety and adherence to ethical guidelines.
Interventions
GENETICStem cell
Primary cell cultures of tissue cells will be established. Cell cultures will undergo genetic reprogramming to induce the long-term propagation of living cells.
GENETICGenetic Screening
Serum samples are processed through RNA sequencing to reveal known and novel infertility-related biomarkers and genes.
GENETICGenetic Reprogramming
Genes or gene products will be reinserted into cells to observe how the cells can be changed, or reprogrammed, into embryonic-like cells or into sperm precursor cells.
GENETICCell Maturation
Genetically unmodified and modified cells are placed in a laboratory-based testicular environment to promote spermatogenesis into maturity.
Sponsors
The Turek Clinic, Inc
CellARTs Inc.
Study design
Allocation
NON_RANDOMIZED
Intervention model
SINGLE_GROUP
Primary purpose
OTHER
Masking
NONE
Intervention model description
All participants will be donating human tissue serum (blood), skin (via biopsy), and testicular tissue (via biopsy). These samples will be used for: 1. Primary cell culture of tissue cells will be established and genetic reprogramming could be applied to induce the long-term propagation of living cells. 2. Characterization and genetic screening of cells will be done to discover and reveal known or novel infertility-related biomarkers or genes. 3. Genetic reprogramming. Certain genes or gene products are inserted into donated cells to study how the cells can be changed, or reprogrammed, into embryonic-like cells or into sperm precursor cells. 4. Culture of cells in testicular organoids. Genetically unmodified or modified cells from donated samples could be placed in a laboratory-based testicular environment to see if sperm can be made.
Eligibility
Sex/Gender
MALE
Age
18 Years to 60 Years
Healthy volunteers
Yes
Inclusion criteria
Group 1 (Fertile Control) * Male sex of reproductive age (between 18 - 60 years old). * Evidence of fertility or normal spermatogenesis. * Will undergo study procedures in conjunction with their planned fertility or infertility procedures performed for clinical purposes. Inclusion Criteria: Group 2 and 3 (Infertile) * Male sex of reproductive age (between 18 - 60 years old). * Males with evidence of \>1 year of infertility. * Posesses diagnosis of azoospermia is on clinical evaluation. * Will undergo study procedures in conjunction with their planned fertility or infertility procedures performed for clinical purposes.
Exclusion criteria
\- The lack of diagnosis of fertility or infertility, and lack of testicles.
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Successful derivation of patient-specific human spermatogonial stem cells (hSSCs) from testicular tissue samples. | From initial sample collection to 24 months post all subject sample collection completion | Metric: The presence of viable hSSCs characterized by specific molecular and cellular markers (e.g., GFRα1, PLZF) within a predefined timeframe post-derivation. Assessment Method: Flow cytometry, immunohistochemistry, or RT-PCR to confirm marker expression. |
Other
| Measure | Time frame | Description |
|---|---|---|
| Development and functional validation of an ex vivo testis organ-on-a-chip ('iTestis') platform for the cultivation and maintenance of isolated spermatogonial stem cells (hSSCs) | From the date of the first successful derivation of patient-specific hSSCs until an ex vivo testis platform is developed and can functionally cultivate and maintain the isolated spermatogonial stem cells (hSSCs), assessed for up to 24 months. | Metric: Successful development of a biomimetic iTestis capable of maintaining hSSCs in a viable and undifferentiated state over a predefined culture period. Assessment Method: Viability: Measured by live/dead assays or metabolic activity (e.g., MTT or ATP assays). Stemness: Expression of hSSC markers (e.g., GFRα1, PLZF) using immunofluorescence or qRT-PCR. |
| Promotion of human spermatogenesis in vitro ('iSperm') using hSSCs and hiPSCs within the iTestis platform. | From the date of the initial development of the ex vivo testis platform until the promotion of human spermatogenesis using hSSCs and hiPSCs is achieved within the iTestis platform, assessed for up to 24 months. | Metric: Generation of haploid spermatogenic cells (e.g., spermatocytes, spermatids, or sperm-like cells) from hSSCs and/or hiPSCs cultured within the iTestis platform. Assessment Method: Flow cytometry or fluorescence-activated cell sorting (FACS) to confirm haploid cell production (e.g., 1N DNA content). Gene and protein expression analysis of spermatogenic markers (e.g., SCP3, acrosin, protamine 1) using RT-PCR and immunofluorescence. |
| Successful derivation of patient-specific induced pluripotent stem cells (hiPSCs) from skin samples. | From initial sample collection to 24 months post all subject sample collection completion. | Metric: Generation of hiPSCs that exhibit hallmark pluripotency characteristics from participant skin fibroblasts. Assessment Method: Morphological Analysis: Observation of colony morphology consistent with hiPSCs. Molecular and Functional Validation: Expression of key pluripotency markers (e.g., OCT4, SOX2, NANOG) verified by immunofluorescence, qRT-PCR, or flow cytometry. Functional pluripotency confirmed via in vitro differentiation into the three germ layers (endoderm, mesoderm, ectoderm). |
Countries
United States
Contacts
Primary ContactErica S Godart, BS
Backup ContactConstance John, PhD
Outcome results
None listed