[Decellularized scaffolds derived from porcine testes for culture of primary testicular cells and organoids].

Decellularized extracellular matrix can well preserve the biomimetic ultrastructure of native tissue, providing a suitable three-dimensional biomimetic microenvironment for the in vitro culture of cells and organoids. The aim of this study is to prepare a porcine testicular decellularized extracellular matrix (PT-dECM) scaffold that fully preserves the natural ultrastructure and bioactive components, thereby providing a highly biomimetic three-dimensional microenvironment for the adhesion and proliferation of primary testicular cells and the in vitro construction of testicular organoids. In this study, we optimized a combined physicochemical decellularization protocol and, after screening, obtained a PT-dECM scaffold that could effectively remove immunogenic substances and fully preserve the natural ultrastructure and bioactive components. This scaffold not only supported uniform cell adhesion and directed proliferation but also provided primary testicular cells with a three-dimensional biomimetic environment that maintained structural and functional integrity. Testicular organoids constructed on this scaffold exhibited the typical physiological structure of native testes, and the four core functional cell types-Sertoli cells, Leydig cells, germ cells, and peritubular myoid cells-were reliably detected, with their spatial distribution being highly consistent with that of native tissue. The PT-dECM scaffold optimized in this study mimicked the natural testicular microenvironment, overcoming the limitations of existing models. It provides a new tool for advancing research into the functions of the testicular microenvironment and has significant application potential in the field of reproductive tissue engineering.