شنبه 10 آبان 1404
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دوره 19، شماره 5 - ( 3-1400 )
جلد 19 شماره 5 صفحات 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-fa.html
URL: http://ijrm.ir/article-1-2930-fa.html
P-45 In vitro mouse spermatogenesis on artificial testis engineered by 3D printing of extracellular matrix. International Journal of Reproductive BioMedicine. 1400; 19 (5) :212-212
چکیده: (394 مشاهده)
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.
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