دورية أكاديمية
Spermatogenesis in mouse testicular organoids with testis-specific architecture, improved germ cell survival and testosterone production.
| العنوان: | Spermatogenesis in mouse testicular organoids with testis-specific architecture, improved germ cell survival and testosterone production. |
|---|---|
| المؤلفون: | Richer G; Biology of the Testis (BITE) laboratory, Genetics Reproduction and Development (GRAD) research group, Vrije Universiteit Brussel (VUB), Brussels, Belgium., Goyvaerts C; Laboratory for Molecular and Cellular Therapy (LMCT), Translational Oncology Research Center (TORC), VUB, Brussels, Belgium.; Department of Medical Imaging, Molecular Imaging and Therapy (MITH), VUB, Brussels, Belgium., Marchandise L; Department of In Vitro Toxicology and Dermato-Cosmetology (IVTD), Center for Pharmaceutical Research, VUB, Brussels, Belgium., Vanhaecke T; Department of In Vitro Toxicology and Dermato-Cosmetology (IVTD), Center for Pharmaceutical Research, VUB, Brussels, Belgium., Goossens E; Biology of the Testis (BITE) laboratory, Genetics Reproduction and Development (GRAD) research group, Vrije Universiteit Brussel (VUB), Brussels, Belgium., Baert Y; Biology of the Testis (BITE) laboratory, Genetics Reproduction and Development (GRAD) research group, Vrije Universiteit Brussel (VUB), Brussels, Belgium.; Department of In Vitro Toxicology and Dermato-Cosmetology (IVTD), Center for Pharmaceutical Research, VUB, Brussels, Belgium. |
| المصدر: | Biofabrication [Biofabrication] 2024 Aug 14; Vol. 16 (4). Date of Electronic Publication: 2024 Aug 14. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: IOP Pub Country of Publication: England NLM ID: 101521964 Publication Model: Electronic Cited Medium: Internet ISSN: 1758-5090 (Electronic) Linking ISSN: 17585082 NLM ISO Abbreviation: Biofabrication Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Bristol : IOP Pub., 2009- |
| مواضيع طبية MeSH: | Organoids*/cytology , Organoids*/metabolism , Organoids*/drug effects , Testis*/cytology , Testis*/drug effects , Testis*/metabolism , Testosterone*/pharmacology , Spermatogenesis*/drug effects , Mice, Inbred C57BL* , Cell Survival*/drug effects, Male ; Animals ; Mice ; Cell Differentiation/drug effects ; Germ Cells/cytology ; Germ Cells/drug effects ; Germ Cells/metabolism ; Dexamethasone/pharmacology |
| مستخلص: | This study presents a biphasic approach to overcome the limitations of current testicular organoid (TO) cultures, including histological heterogeneity, germ cell loss and absence of spermatogenesis. Agarose microwells were utilized to create TOs from prepubertal C57BL/6 J testicular cells. First emphasis was on improving germ cell survival during the initial 2-week reorganization phase by comparing α -MEM + 10% knockout serum replacement (KSR) medium, known to support TO generation in mice, to three optimized media (1-3). Cell densities and culture dynamics were also tested to recreate histological resemblance to testes. After optimizing germ cell survival and cell organization, the effect of growth factors and immunomodulation through CD45 + immune cell depletion or dexamethasone (DEX) supplementation were assessed for enhancing spermatogenesis during the subsequent differentiation phase. Testicular cells self-reorganized into organoids resembling the testicular anatomical unit, characterized by one tubule-like structure surrounded by interstitium. Media 1-3 proved superior for organoid growth during the reorganization phase, with TOs in medium 3 exhibiting germ cell numbers (7.4% ± 4.8%) comparable to controls (9.3% ± 5.3%). Additionally, 37% ± 30% demonstrated organized histology from 32 × 10 3 cells under static conditions. Switching to α -MEM + 10% KSR during the differentiation phase increased formation efficiency to 85 ± 7%, along with elevated germ cell numbers, testosterone production (3.1 ± 0.9 ng ml -1 ) and generation of γ- H2AX + spermatid-like cells (steps 8-11, 1.2% ± 2.2% of the total). Adding differentiation factors to the α -MEM increased spermatid-like cell numbers to 2.9% ± 5.9%, confirmed through positive staining for CREM, transition protein 1, and peanut agglutinin. Although, these remained diploid with irregular nuclear maturation. DEX supplementation had no additional effect, and immune cell depletion adversely impacted TO formation. The manipulability of TOs offers advantages in studying male infertility and exploring therapies, with scalability enabling high-throughput chemical screening and reducing animal usage in reproductive toxicity and drug discovery studies. (Creative Commons Attribution license.) |
| فهرسة مساهمة: | Keywords: in vitro spermatogenesis; male infertility; reproductive toxicity; spermatogonial stem cells; testicular organoids |
| المشرفين على المادة: | 3XMK78S47O (Testosterone) 7S5I7G3JQL (Dexamethasone) |
| تواريخ الأحداث: | Date Created: 20240710 Date Completed: 20240814 Latest Revision: 20240814 |
| رمز التحديث: | 20240814 |
| DOI: | 10.1088/1758-5090/ad618f |
| PMID: | 38986466 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1758-5090 |
|---|---|
| DOI: | 10.1088/1758-5090/ad618f |