عنوان	Engineering of Chitosan Hydrogel Covered with Testicular Cell’s Extracellular Matrix as An Excellent Substrate for Spermatogonial Stem Cells Adhesion and Proliferation
نوع پژوهش	مقاله ارائه‌شده
کلیدواژه‌ها	Keywords: Hydrogel Systems, Cell Scaffold, Testicular Cells, Extracellular Matrix, Spermatogonia Stem Cells
چکیده	Background: Successful in vitro transplantation of spermatogonial stem cells (SSCs) requires efficient culture systems. To mimic the natural environment, this study used a temperaturesensitive chitosan-based hydrogel scaffold coated with testicular cell-derived extracellular matrix. This scaffold supported the co-culture of testicular and spermatogonial cells, promoting SSC proliferation and differentiation into sperm-producing cells. Materials and Methods: In this study, a chitosan-based thermos- responsive hydrogel was designed and its morphology and gelation time were investigated. Testicular cells from NMRI mice were cultured on the hydrogel for 14 days to deposit extracellular matrix, after which the scaffolds and various tests of morphology were decellularized. The effect of ECM-coated hydrogel (D-Hydrogel) on spermatogonial stem cell reproductive gene expression was evaluated using RT-PCR and compared with Hydrogel and control groups. Results: DAPI staining confirmed the absence of cells and debris in the extracellular matrix (ECM). Coating the hydrogel with ECM increased water absorption and degradation (P<0.001). DNA analysis showed significantly higher genetic content in the TC-Hydrogel group compared to Hydrogel and D-Hydrogel groups (P<0.001). SSC survival was significantly higher on ECM-coated hydrogel (D-Hydrogel) than on uncoated hydrogel (P<0.001). Cells on D-Hydrogel showed enhanced adhesion and elongation. Molecular analysis revealed a significant increase in SSC proliferation gene expression in the D-Hydrogel group compared to Hydrogel and control at days 7 (P≤0.05) and 14 (P<0.001). Conclusion: This study introduces thermo-sensitive hydrogel scaffolds coated with extracellular matrix of testicular cells as a suitable system for culturing and proliferating spermatogonia stem cells and possibly a suitable method for sperm production in vitro. However, further studies are needed to determine the effect of D-Hydrogel on differentiation of spermatogonia cells to produce sperm in the future studies.
پژوهشگران	منیره محمودی (نفر دوم)، مهدیه قلی پور ملک آبادی (نفر سوم)، مهشاد کاهدی (نفر اول)