Document Type : Original Article

Authors

1 Fertility and Infertility Research Center, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran

2 Student Research Committee, Kermanshah University of Medical Sciences, Kermanshah, Iran

3 Department of Tissue Engineering, School of Medicine, Kermanshah University of Medical Sciences, Kermanshah, Iran

Abstract

Background: Men’s infertility and lack of production of healthy and active sperm are concerns of recent years in most
countries. Studies on the preparation of extracellular matrix (ECM) from decellularization of testis tissue and spermatogenesis
could provide proper results to solve some of the men’s infertility 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 were decellularized by a 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 analyzed 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: Methods of 1% SDS, 1% SDS-vacuum, and 1% SD completely removed the cells. The preservation of collagen
and GAG was confirmed using the staining kit and methods. The use of a vacuum showed greater porosity in
the 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 ECM
structure to a large extent without having cytotoxic and hemolysis effects.

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