Document Type : Original Article
Fertility and Infertility Research Center, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran
Student Research Committee, Kermanshah University of Medical Sciences, Kermanshah, Iran
Background: Men's infertility and lack of production of healthy and active sperm are concerns of recent years in most countries Preparation of extracellular matrix (ECM) from decellularization of testis tissue and spermatogenesis studies is a suitable solution to solve some problems. This study aims to decellularize calf testis by different methods to reach a suitable scaffold and introduce it in spermatogenesis studies.
Materials and methods: In this experimental study, calf testis was decellularized by freeze-de freeze, 1% sodium deoxycholate (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 tested using the kit and staining with Hematoxylin-Eosin, Masson's trichrome, Alcian blue, and Orcein methods. The morphology of the scaffolds was analyzed with a scanning electron microscope (SEM).
Results: Biocompatibility and hemocompatibility of the scaffolds were measured. Methods of 1% SDS, 1% SDS- vacuum, and 1%SD completely removed the cells. Preservation of collagen and GAG was confirmed by staining kit and methods. The use of a vacuum showed greater porosity in the SEM images. Toxicity and hemolysis were not seen in the scaffolds.
Conclusion: Testis decellularization with 1%SDS and 1%SD, in addition to cell removal, can maintain the ECM structure to a large extent without having cytotoxic and hemolysis effects.