Localization and characterization of SSCs from pre-pubertal bovine testes

MOHAMMAD KAZIM WARDAK; KAUSHALYA KULATHUNGA; CHATHURA PRIYADARSHANA

doi:10.56093/ijans.v92i10.124617

Authors

MOHAMMAD KAZIM WARDAK Department of Animal Science, Faculty of Agricultural Science, Shaikhzayed University, Khost 2501, Afghanistan
KAUSHALYA KULATHUNGA Department of Physiology, Faculty of Medicine, Sabaragamuwa University of Sri Lanka, No. 01, Hidallena, Ratnapura, Sri Lanka
CHATHURA PRIYADARSHANA Animal Health Division, Hayleys Agriculture Holdings Limited, No. 25, Foster Lane, Colombo 10, Sri Lanka

https://doi.org/10.56093/ijans.v92i10.124617

Keywords:

Bovine, Characterization, Localization, Pre-pubertal, SSCs, Testis

Abstract

Self renewal and proliferation ability of spermatogonial stem cells (SSCs) support spermatogenesis during adult life. Theoretically, these stem cells can be utilized for transmission of genetic information to descendants via testicular transplantation. However, lack of knowledge in methodologies for identification of SSCs limits the application of SSCs transplantation in domestic animals. Accumulated studies have shown that SSCs specific markers (DBA, UCHL1) and stem cell marker (Sox2, Oct4) are useful to screen SSCs that able to be used for transplantation. However, in cattle, less information is available on the expression status of these markers till date. Therefore, a study was carried out in 2019 at Tsukuba University, Japan where testes from 3, 5 and 7 months old calves were utilized to examine testicular localization and in vitro propogation of stem cell markers. SSCs were isolated by enzymatic digestion combined with centrifugal separation on discontinuous Percoll density gradient. Cell propagation and SSCs marker expression were determined at 5, 10 and 15 days post-culture. Immunostaining in conjunction with Western Blot analysis of cultured cells showed that stem cell markers (UCHL1, Oct4 and Sox2) were expressed in SSCs suggesting that differentiation of gonocyte started by 3 months and SSCs differentiation begins after 5 months of age. Taken together, these results demonstrated marker expression and localization of bull SSCs and showed that in vitro culturing of bull SSCs is implementable.

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Author Biography

CHATHURA PRIYADARSHANA, Animal Health Division, Hayleys Agriculture Holdings Limited, No. 25, Foster Lane, Colombo 10, Sri Lanka

Manager - Animal Health Division

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