The Decellularized Calf Testis: Introducing Suitable Scaffolds for Spermatogenesis Studies

Abstract

Background: Men’s infertility and lack of production of healthy and active sperm are concerns of recent years in mostcountries. Studies on the preparation of extracellular matrix (ECM) from decellularization of testis tissue and spermatogenesiscould provide proper results to solve some of the men’s infertility problems. This study aims to decellularize calftestis by different methods to reach a suitable scaffold and introduce it in spermatogenesis studies.Materials and Methods: In this experimental study, calf testis were decellularized by a freeze-de freeze, 1% sodiumdeoxycholate (SD), 0.1% sodium dodecyl sulfate (SDS), 0.1% SDS-vacuum, 1% SDS, 1% SDS-vacuum, and Triton-X100 methods. The content of DNA, collagen, and glycosaminoglycan (GAG) was analyzed using the kit and stainingwith Hematoxylin-Eosin, Masson’s trichrome, Alcian blue, and Orcein methods. The morphology of the scaffolds wasanalyzed with a scanning electron microscope (SEM).Results: Methods of 1% SDS, 1% SDS-vacuum, and 1% SD completely removed the cells. The preservation of collagenand GAG was confirmed using the staining kit and methods. The use of a vacuum showed greater porosity inthe SEM images. Toxicity and hemolysis were not observed in the scaffolds.Conclusion: Testis decellularization with 1% SDS and 1% SD, in addition to cell removal, could maintain the ECMstructure to a large extent without having cytotoxic and hemolysis effects.

Keywords

• decellularization
• scaffold
• testis