LAUSR

In vitro production of sperm is a desirable idea for fertility preservation in azoospermic men and prepubertal boys suffering from cancer. In this study, a biocompatible porous scaffold based on a triad mixture of silk fibroin (SF), alginate (Alg) and laminin (LM) was developed to facilitate the differentiation of mouse spermatogonia stem cells (SSCs). Following SF extraction, the content was analyzed by SDS-PAGE and stable porous 3D scaffolds were successfully prepared by merely Alg, SF, and a combination of Alg-SF, or Alg-SF-LM through freeze-drying. Then, the biomimetic scaffolds were characterized regarding the structural and biological properties, water absorption capacity, biocompatibility, biodegradability and mechanical behavior. Neonatal mice testicular cells were seeded on 3D scaffolds and following characterization steps, their differentiation efficiency was evaluated using real-time PCR, flow cytometry, immunohistochemistry and H&E staining. Blend matrices showed uniform porous microstructures with interconnected networks, which maintained long-term stability and mechanical properties better than homogenous structures. Molecular analysis of the cells after 21 days of culture showed that the expression of differentiation-related proteins in cells that were developed in composite scaffolds was significantly higher than in other groups. The application of a composite system can lead to the differentiation of SSCs, paving the way for a novel infertility treatment landscape in the future. This article is protected by copyright. All rights reserved.