Immature rat testis sustained long-term development using an integrative model
Abstract Background Xenotransplantation has been primarily performed using fresh donor tissue to study testicular development for about 20 years, and whether the cultured tissue would be a suitable donor is unclear. In this study, we combined testicular culture and xenotransplantation into an integr...
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| Format: | Article |
| Language: | English |
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BMC
2022-10-01
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| Series: | Biological Research |
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| Online Access: | https://doi.org/10.1186/s40659-022-00398-y |
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| author | Yubo Ma Juan Chen Hecheng Li Fangshi Xu Tie Chong Ziming Wang Liandong Zhang |
| author_facet | Yubo Ma Juan Chen Hecheng Li Fangshi Xu Tie Chong Ziming Wang Liandong Zhang |
| author_sort | Yubo Ma |
| collection | DOAJ |
| description | Abstract Background Xenotransplantation has been primarily performed using fresh donor tissue to study testicular development for about 20 years, and whether the cultured tissue would be a suitable donor is unclear. In this study, we combined testicular culture and xenotransplantation into an integrative model and explored whether immature testicular tissue would survive and continue to develop in this model. Methods In the new integrative model group, the testes of neonatal rats on postnatal day 8 (PND 8) were cultured for 4 days ex vivo and then were transplanted under the dorsal skin of castrated nude mice. The xenografted testes were resected on the 57th day after xenotransplantation and the testes of rats in the control group were harvested on PND 69. The survival state of testicular tissue was evaluated from morphological and functional perspectives including H&E staining, immunohistochemical staining of 8-OH-dG, immunofluorescence staining, TUNEL assay, ultrastructural study, gene expression and protein analysis. Results (a) We found that complete spermatogenesis was established in the testes in the new integrative model group. Compared with the control in the same stage, the seminiferous epithelium in some tubules was a bit thinner and there were vacuoles in part of the tubules. Immunofluorescence staining revealed some ACROSIN-positive spermatids were present in seminiferous tubule of xenografted testes. TUNEL detection showed apoptotic cells and most of them were germ cells in the new integrative model group. 8-OH-dG immunohistochemistry showed strongly positive-stained in the seminiferous epithelium after xenotransplantation in comparison with the control group; (b) Compared with the control group, the expressions of FOXA3, DAZL, GFRα1, BOLL, SYCP3, CDC25A, LDHC, CREM and MKI67 in the new integrative model group were significantly elevated (P < 0.05), indicating that the testicular tissue was in an active differentiated and proliferative state; (c) Antioxidant gene detection showed that the expression of Nrf2, Keap1, NQO1 and SOD1 in the new integrative model group was significantly higher than those in the control group (P < 0.05), and DNA methyltransferase gene detection showed that the expression of DNMT3B was significantly elevated as well (P < 0.05). Conclusion The new integrative model could maintain the viability of immature testicular tissue and sustain the long-term survival in vivo with complete spermatogenesis. However, testicular genes expression was altered, vacuolation and thin seminiferous epithelium were still apparent in this model, manifesting that oxidative damage may contribute to the testicular development lesion and it needs further study in order to optimize this model. |
| first_indexed | 2024-04-12T00:32:41Z |
| format | Article |
| id | doaj.art-0332f5e488524ed19ca04235f21e8e63 |
| institution | Directory Open Access Journal |
| issn | 0717-6287 |
| language | English |
| last_indexed | 2024-04-12T00:32:41Z |
| publishDate | 2022-10-01 |
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| spelling | doaj.art-0332f5e488524ed19ca04235f21e8e632022-12-22T03:55:17ZengBMCBiological Research0717-62872022-10-0155111310.1186/s40659-022-00398-yImmature rat testis sustained long-term development using an integrative modelYubo Ma0Juan Chen1Hecheng Li2Fangshi Xu3Tie Chong4Ziming Wang5Liandong Zhang6Department of Urology, The Second Affiliated Hospital of Xi’an Jiaotong UniversityDepartment of Urology, The Second Affiliated Hospital of Xi’an Jiaotong UniversityDepartment of Urology, The Second Affiliated Hospital of Xi’an Jiaotong UniversityDepartment of Urology, The Second Affiliated Hospital of Xi’an Jiaotong UniversityDepartment of Urology, The Second Affiliated Hospital of Xi’an Jiaotong UniversityDepartment of Urology, The Second Affiliated Hospital of Xi’an Jiaotong UniversityDepartment of Urology, The Second Affiliated Hospital of Xi’an Jiaotong UniversityAbstract Background Xenotransplantation has been primarily performed using fresh donor tissue to study testicular development for about 20 years, and whether the cultured tissue would be a suitable donor is unclear. In this study, we combined testicular culture and xenotransplantation into an integrative model and explored whether immature testicular tissue would survive and continue to develop in this model. Methods In the new integrative model group, the testes of neonatal rats on postnatal day 8 (PND 8) were cultured for 4 days ex vivo and then were transplanted under the dorsal skin of castrated nude mice. The xenografted testes were resected on the 57th day after xenotransplantation and the testes of rats in the control group were harvested on PND 69. The survival state of testicular tissue was evaluated from morphological and functional perspectives including H&E staining, immunohistochemical staining of 8-OH-dG, immunofluorescence staining, TUNEL assay, ultrastructural study, gene expression and protein analysis. Results (a) We found that complete spermatogenesis was established in the testes in the new integrative model group. Compared with the control in the same stage, the seminiferous epithelium in some tubules was a bit thinner and there were vacuoles in part of the tubules. Immunofluorescence staining revealed some ACROSIN-positive spermatids were present in seminiferous tubule of xenografted testes. TUNEL detection showed apoptotic cells and most of them were germ cells in the new integrative model group. 8-OH-dG immunohistochemistry showed strongly positive-stained in the seminiferous epithelium after xenotransplantation in comparison with the control group; (b) Compared with the control group, the expressions of FOXA3, DAZL, GFRα1, BOLL, SYCP3, CDC25A, LDHC, CREM and MKI67 in the new integrative model group were significantly elevated (P < 0.05), indicating that the testicular tissue was in an active differentiated and proliferative state; (c) Antioxidant gene detection showed that the expression of Nrf2, Keap1, NQO1 and SOD1 in the new integrative model group was significantly higher than those in the control group (P < 0.05), and DNA methyltransferase gene detection showed that the expression of DNMT3B was significantly elevated as well (P < 0.05). Conclusion The new integrative model could maintain the viability of immature testicular tissue and sustain the long-term survival in vivo with complete spermatogenesis. However, testicular genes expression was altered, vacuolation and thin seminiferous epithelium were still apparent in this model, manifesting that oxidative damage may contribute to the testicular development lesion and it needs further study in order to optimize this model.https://doi.org/10.1186/s40659-022-00398-yNeonatal testisOrgan cultureXenotransplantationSpermatogenesisOxidative stress |
| spellingShingle | Yubo Ma Juan Chen Hecheng Li Fangshi Xu Tie Chong Ziming Wang Liandong Zhang Immature rat testis sustained long-term development using an integrative model Biological Research Neonatal testis Organ culture Xenotransplantation Spermatogenesis Oxidative stress |
| title | Immature rat testis sustained long-term development using an integrative model |
| title_full | Immature rat testis sustained long-term development using an integrative model |
| title_fullStr | Immature rat testis sustained long-term development using an integrative model |
| title_full_unstemmed | Immature rat testis sustained long-term development using an integrative model |
| title_short | Immature rat testis sustained long-term development using an integrative model |
| title_sort | immature rat testis sustained long term development using an integrative model |
| topic | Neonatal testis Organ culture Xenotransplantation Spermatogenesis Oxidative stress |
| url | https://doi.org/10.1186/s40659-022-00398-y |
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