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International Journal of Reproductive BioMedicine
Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences of Yazd
ISSN: 1680-6433 EISSN: 1680-6433
Vol. 19, No. 4, 2021, pp. 321-332
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Bioline Code: rm21030
Full paper language: English
Document type: Research Article
Document available free of charge
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International Journal of Reproductive BioMedicine, Vol. 19, No. 4, 2021, pp. 321-332
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Comparison of two methods for prolong storage of decellularized mouse whole testis for tissue engineering application: An experimental study
Majidi Gharenaz, Nasrin; Movahedin, Mansoureh & Mazaheri, Zohreh
Abstract
Background: Biological scaffolds are derived by the decellularization of tissues or
organs. Various biological scaffolds, such as scaffolds for the liver, lung, esophagus,
dermis, and human testicles, have been produced. Their application in tissue
engineering has created the need for cryopreservation processes to store these
scaffolds.
Objective: The aim was to compare the two methods for prolong storage testicular
scaffolds.
Materials and Methods: In this experimental study, 20 male NMRI mice (8 wk) were
sacrificed and their testes were removed and treated with 0.5% sodium dodecyl sulfate
followed by Triton X-100 0.5%. The efficiency of decellularization was determined
by histology and DNA quantification. Testicular scaffolds were stored in phosphate-
buffered saline solution at 4°C or cryopreserved by programmed slow freezing followed
by storage in liquid nitrogen. Masson’s trichrome staining, Alcian blue staining and
immunohistochemistry, collagen assay, and glycosaminoglycan assay were done prior
to and after six months of storage under each condition.
Results: Hematoxylin-eosin staining showed no remnant cells after the completion
of decellularization. DNA content analysis indicated that approximately 98% of the
DNA was removed from the tissue (p = 0.02). Histological evaluation confirmed
the preservation of extracellular matrix components in the fresh and frozen-thawed
scaffolds. Extracellular matrix components were decreased by 4°C-stored scaffolds.
Cytotoxicity tests with mouse embryonic fibroblast showed that the scaffolds were
biocompatible and did not have a harmful effect on the proliferation of mouse
embryonic fibroblast cells.
Conclusion: Our results demonstrated the superiority of the slow freezing method for
prolong storage of testicular scaffolds.
Keywords
Cryopreservation; Testis; Scaffold; Mouse.
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© Copyright 2021 - Majidi Gharenaz et al. Alternative site location: http://www.ijrm.ir
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