International Journal of
Reproductive Biomedicine
Wed, May 14, 2025
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Volume 19, Issue 5 (Suppl- 2021)
IJRM 2021, 19(5): 212-212 |
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Bashiri Nahangi Z, Amiri I, Gholipourmalekabadi M, Falak R, Asgari H, Koruji M. P-45 In vitro mouse spermatogenesis on artificial testis engineered by 3D printing of extracellular matrix. IJRM 2021; 19 (5) :212-212
URL: http://ijrm.ir/article-1-2930-en.html
URL: http://ijrm.ir/article-1-2930-en.html
1- Stem Cell and Regenerative Medicine Research Center, Iran University of Medical Sciences, Tehran, Iran Department of Anatomy, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
2- Research Center for Molecular Medicine, Hamadan University of Medical Sciences, Hamadan, Iran Endometrium and Research Center, Hamadan University of Medical Sciences, Hamadan, Iran
3- Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, Iran Department of Tissue Engineering and Regenerative Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran
4- Immunology Research Center (IRC), Institute of Immunology and Infectious Diseases, Iran University of Medical Sciences, Tehran, Iran
5- Stem Cell and Regenerative Medicine Research Center, Iran University of Medical Sciences, Tehran, Iran Department of Anatomy, School of Medicine, Iran University of Medical Sciences, Tehran, Iran ,koruji.m@iums.ac.ir
2- Research Center for Molecular Medicine, Hamadan University of Medical Sciences, Hamadan, Iran Endometrium and Research Center, Hamadan University of Medical Sciences, Hamadan, Iran
3- Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, Iran Department of Tissue Engineering and Regenerative Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran
4- Immunology Research Center (IRC), Institute of Immunology and Infectious Diseases, Iran University of Medical Sciences, Tehran, Iran
5- Stem Cell and Regenerative Medicine Research Center, Iran University of Medical Sciences, Tehran, Iran Department of Anatomy, School of Medicine, Iran University of Medical Sciences, Tehran, Iran ,
Abstract: (181 Views)
Background: Male infertility accounts for about 50% of all infertility cases, and 25 % of infertile men are azoospermic. Due to the very small number of spermatogonia stem cells (SSCs) in testicular tissue biopsy specimens, SSCs culture for infertile patients can be important.
Objective: The proliferation of SSCs on printed scaffold derived from the extracellular matrix (ECM) of testicular tissues evaluated.
Materials and Methods: Ram testicular tissue was decellularized using hypertonic solution -Triton X-100 for 30 min. The extracted ECM (5% ratio) was used as a bio-ink for the fabrication of artificial testes along with alginate and gelatin. Testicular cells were then isolated from the testes of 3-7 days old neonate mice after enzymatic digestion. The nature of SSCs was confirmed by flow cytometry and RT-PCR for specific markers Plzf, Id4, Gfrα1, and Prm1. Finally, cell viability evaluated using MTT test and testicular cell proliferation process on printed alginate-gelatin scaffolds (group I) and ECM-alginate-gelatin scaffolds (group II) using immunocytochemistry, flow cytometry, and real-time PCR techniques was assessed.
Results: The MTT test indicated that the cell viability on the composite scaffold was significantly higher than the hybrid scaffolds and control group (p > 0.05). The results of 2 wk of proliferation on the printed system showed that the expression of Plzf, Id4, Gfrα1 gene using real-time PCR in group II was significantly higher than group I (p > 0.05). Flow cytometry analysis also showed that the number of Plzf-positive cells in group II was significantly higher than group I (p > 0.05). Immunocytochemistry results confirmed that Plzf, Id4, and Gfrα1 markers were expressed in both groups, but their expression in group II was significantly higher than group I (p > 0.05).
Conclusion: We concluded that the culture of testicular cells on scaffolds containing ECM increases the viability, colonization, and proliferation of SSCs and achieves a high number of cells for differentiation in vitro. Therefore, 3D printing using the ECM of the testis can be an ideal strategy for the regeneration of seminiferous tubules.
Objective: The proliferation of SSCs on printed scaffold derived from the extracellular matrix (ECM) of testicular tissues evaluated.
Materials and Methods: Ram testicular tissue was decellularized using hypertonic solution -Triton X-100 for 30 min. The extracted ECM (5% ratio) was used as a bio-ink for the fabrication of artificial testes along with alginate and gelatin. Testicular cells were then isolated from the testes of 3-7 days old neonate mice after enzymatic digestion. The nature of SSCs was confirmed by flow cytometry and RT-PCR for specific markers Plzf, Id4, Gfrα1, and Prm1. Finally, cell viability evaluated using MTT test and testicular cell proliferation process on printed alginate-gelatin scaffolds (group I) and ECM-alginate-gelatin scaffolds (group II) using immunocytochemistry, flow cytometry, and real-time PCR techniques was assessed.
Results: The MTT test indicated that the cell viability on the composite scaffold was significantly higher than the hybrid scaffolds and control group (p > 0.05). The results of 2 wk of proliferation on the printed system showed that the expression of Plzf, Id4, Gfrα1 gene using real-time PCR in group II was significantly higher than group I (p > 0.05). Flow cytometry analysis also showed that the number of Plzf-positive cells in group II was significantly higher than group I (p > 0.05). Immunocytochemistry results confirmed that Plzf, Id4, and Gfrα1 markers were expressed in both groups, but their expression in group II was significantly higher than group I (p > 0.05).
Conclusion: We concluded that the culture of testicular cells on scaffolds containing ECM increases the viability, colonization, and proliferation of SSCs and achieves a high number of cells for differentiation in vitro. Therefore, 3D printing using the ECM of the testis can be an ideal strategy for the regeneration of seminiferous tubules.
Type of Study: Congress Abstract |
Subject:
Reproductive Pattology
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