Effectiveness of Voytik-Harbin Protocol in Fabrication of Ram’s Testicular-Derived Hydrogel and Its Impact on Mouse In Vitro Spermatogenesis
Background: The utilization of decellularized extracellular matrix (dECM) derived from animal testis tissue hasdemonstrated potential as a component of tissue-specific scaffolds. Current research is mostly centered arounddECM as a natural resource for culturing testicular cells. This study aimed to...
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Royan Institute (ACECR), Tehran
2025-01-01
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| Series: | International Journal of Fertility and Sterility |
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| Online Access: | https://www.ijfs.ir/article_712722_8c275281154eebcc3731c14411a7f7d5.pdf |
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| author | Leila Rashki Ghaleno Tohid Rezaei Topraggaleh Leila Montazeri Abdolhossein Shahverdi Mojtaba Rezazadeh Valojerdi |
| author_facet | Leila Rashki Ghaleno Tohid Rezaei Topraggaleh Leila Montazeri Abdolhossein Shahverdi Mojtaba Rezazadeh Valojerdi |
| author_sort | Leila Rashki Ghaleno |
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| description | Background: The utilization of decellularized extracellular matrix (dECM) derived from animal testis tissue hasdemonstrated potential as a component of tissue-specific scaffolds. Current research is mostly centered arounddECM as a natural resource for culturing testicular cells. This study aimed to assess firstly the comparison ofVoytik-Harbin (VH) and Frytes protocol in creating Ram’s dECM testis hydrogel and secondly the evaluation ofthe best protocol effect on in vitro spermatogenesis.Materials and Methods: In this experimental study, the six testes of mature rams were decellularized and the hydrogelproduction was performed by i. The Frytes protocol utilized a concentration of 1 mg/mL of pepsin, dissolvedin either 0.1 or 0.01 M HCl, and ii. The VH protocol was involved 10 mg of pepsin per 100 mg of ECM in 0.5 M ofacetic acid. Subsequently, mouse testicular cells were cultivated on collagen hydrogel as the control and the moreeffective testicular-derived hydrogel (TDH) to evaluate the early stages of in vitro spermatogenesis.Results: While the Freytes protocol produced a homogeneous pre-gel solution with both HCl concentrations; elevatingthe pH to 7.4 loosened the hydrogel and made gelation problematic. In contrast, the VH protocol solidifiedthe hydrogel and produced a strong hydrogel due to its gelation consistency. Furthermore, the prepared hydrogel byVH with 25 mg of dECM had a significantly higher priority in terms of rheology and structure (P<0.05). Followingmouse testicular cell culture, TDH and collagen hydrogel did not differ significantly in terms of cell survival ratesand the mRNA expression of early spermatogenesis genes.Conclusion: Using the VH protocol for producing ram TDH resulted in a firm hydrogel with a high frequency ofrepeat, which may be suited for testicular cell growth. |
| format | Article |
| id | doaj-art-745b9cadc3e345e5a6c2bfcdb0c5f480 |
| institution | Kabale University |
| issn | 2008-076X 2008-0778 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Royan Institute (ACECR), Tehran |
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| series | International Journal of Fertility and Sterility |
| spelling | doaj-art-745b9cadc3e345e5a6c2bfcdb0c5f4802025-01-18T09:23:38ZengRoyan Institute (ACECR), TehranInternational Journal of Fertility and Sterility2008-076X2008-07782025-01-01191707910.22074/ijfs.2024.2018868.1599712722Effectiveness of Voytik-Harbin Protocol in Fabrication of Ram’s Testicular-Derived Hydrogel and Its Impact on Mouse In Vitro SpermatogenesisLeila Rashki Ghaleno0Tohid Rezaei Topraggaleh1Leila Montazeri2Abdolhossein Shahverdi3Mojtaba Rezazadeh Valojerdi4Department of Reproductive Biology, Faculty of Basic Sciences and Advanced Medical Technologies, Royan Institute, ACECR, Tehran, IranReproductive Health Research Center, Clinical Research Institute, Urmia University of Medical Sciences, Urmia, IranDepartment of Cell Engineering, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, IranDepartment of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, IranDepartment of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, IranBackground: The utilization of decellularized extracellular matrix (dECM) derived from animal testis tissue hasdemonstrated potential as a component of tissue-specific scaffolds. Current research is mostly centered arounddECM as a natural resource for culturing testicular cells. This study aimed to assess firstly the comparison ofVoytik-Harbin (VH) and Frytes protocol in creating Ram’s dECM testis hydrogel and secondly the evaluation ofthe best protocol effect on in vitro spermatogenesis.Materials and Methods: In this experimental study, the six testes of mature rams were decellularized and the hydrogelproduction was performed by i. The Frytes protocol utilized a concentration of 1 mg/mL of pepsin, dissolvedin either 0.1 or 0.01 M HCl, and ii. The VH protocol was involved 10 mg of pepsin per 100 mg of ECM in 0.5 M ofacetic acid. Subsequently, mouse testicular cells were cultivated on collagen hydrogel as the control and the moreeffective testicular-derived hydrogel (TDH) to evaluate the early stages of in vitro spermatogenesis.Results: While the Freytes protocol produced a homogeneous pre-gel solution with both HCl concentrations; elevatingthe pH to 7.4 loosened the hydrogel and made gelation problematic. In contrast, the VH protocol solidifiedthe hydrogel and produced a strong hydrogel due to its gelation consistency. Furthermore, the prepared hydrogel byVH with 25 mg of dECM had a significantly higher priority in terms of rheology and structure (P<0.05). Followingmouse testicular cell culture, TDH and collagen hydrogel did not differ significantly in terms of cell survival ratesand the mRNA expression of early spermatogenesis genes.Conclusion: Using the VH protocol for producing ram TDH resulted in a firm hydrogel with a high frequency ofrepeat, which may be suited for testicular cell growth.https://www.ijfs.ir/article_712722_8c275281154eebcc3731c14411a7f7d5.pdfcell culture techniquedecellularized extracellular matrixhydrogelsspermatogenesis |
| spellingShingle | Leila Rashki Ghaleno Tohid Rezaei Topraggaleh Leila Montazeri Abdolhossein Shahverdi Mojtaba Rezazadeh Valojerdi Effectiveness of Voytik-Harbin Protocol in Fabrication of Ram’s Testicular-Derived Hydrogel and Its Impact on Mouse In Vitro Spermatogenesis International Journal of Fertility and Sterility cell culture technique decellularized extracellular matrix hydrogels spermatogenesis |
| title | Effectiveness of Voytik-Harbin Protocol in Fabrication of Ram’s Testicular-Derived Hydrogel and Its Impact on Mouse In Vitro Spermatogenesis |
| title_full | Effectiveness of Voytik-Harbin Protocol in Fabrication of Ram’s Testicular-Derived Hydrogel and Its Impact on Mouse In Vitro Spermatogenesis |
| title_fullStr | Effectiveness of Voytik-Harbin Protocol in Fabrication of Ram’s Testicular-Derived Hydrogel and Its Impact on Mouse In Vitro Spermatogenesis |
| title_full_unstemmed | Effectiveness of Voytik-Harbin Protocol in Fabrication of Ram’s Testicular-Derived Hydrogel and Its Impact on Mouse In Vitro Spermatogenesis |
| title_short | Effectiveness of Voytik-Harbin Protocol in Fabrication of Ram’s Testicular-Derived Hydrogel and Its Impact on Mouse In Vitro Spermatogenesis |
| title_sort | effectiveness of voytik harbin protocol in fabrication of ram s testicular derived hydrogel and its impact on mouse in vitro spermatogenesis |
| topic | cell culture technique decellularized extracellular matrix hydrogels spermatogenesis |
| url | https://www.ijfs.ir/article_712722_8c275281154eebcc3731c14411a7f7d5.pdf |
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