Generation of Organized Porcine Testicular Organoids in Solubilized Hydrogels from Decellularized Extracellular Matrix.
Maxime VermeulenFederico Del VentoMarc KanbarSebastien Pyr Dit RuysDidier VertommenJonathan PoelsChristine WynsPublished in: International journal of molecular sciences (2019)
Cryopreservation of immature testicular tissue (ITT) prior to chemo/radiotherapy is now ethically accepted and is currently the only way to preserve fertility of prepubertal boys about to undergo cancer therapies. So far, three-dimensional culture of testicular cells isolated from prepubertal human testicular tissue was neither efficient nor reproducible to obtain mature spermatozoa, and ITT transplantation is not a safe option when there is a risk of cancer cell contamination of the testis. Hence, generation of testicular organoids (TOs) after cell selection is a novel strategy aimed at restoring fertility in these patients. Here, we created TOs using hydrogels developed from decellularized porcine ITT and compared cell numbers, organization and function to TOs generated in collagen only hydrogel. Organotypic culture of porcine ITT was used as a control. Rheological and mass spectrometry analyses of both hydrogels highlighted differences in terms of extracellular matrix stiffness and composition, respectively. Sertoli cells (SCs) and germ cells (GCs) assembled into seminiferous tubule-like structures delimited by a basement membrane while Leydig cells (LCs) and peritubular cells localized outside. TOs were maintained for 45 days in culture and secreted stem cell factor and testosterone demonstrating functionality of SCs and LCs, respectively. In both TOs GC numbers decreased and SC numbers increased. However, LC numbers decreased significantly in the collagen hydrogel TOs (p < 0.05) suggesting a better preservation of growth factors within TOs developed from decellularized ITT and thus a better potential to restore the reproductive capacity.
Keyphrases
- extracellular matrix
- induced apoptosis
- cell cycle arrest
- mass spectrometry
- tissue engineering
- stem cells
- endoplasmic reticulum stress
- oxidative stress
- germ cell
- end stage renal disease
- squamous cell carcinoma
- risk assessment
- radiation therapy
- newly diagnosed
- ms ms
- high resolution
- cancer therapy
- prognostic factors
- peritoneal dialysis
- childhood cancer
- photodynamic therapy
- locally advanced
- smoking cessation
- health risk