Effect of stage of puberty on culture characteristics of goat spermatogonial stem cells

Abstract

The aim of experiment was to study the culture characteristics of spermatogonial stem cells (SSCs) obtained from pre-and post-pubertal goat testes. Six animals each of two age groups (3-6 months and 1.0-1.5 years) were included in the study. The testes were collected from slaughtered animals and after washing, morpho-biometric characters such as weight, length, mid circumference and volume of both the left and right testes were recorded. The isolated SSCs were enriched by double filtration through 80 and 60 μm nylon mesh filters. The filtrate was cultured overnight with SSC medium Dulbecco's modified eagle's medium / HamF-12 medium. The colonies were also characterized by alkaline phosphatase activity and demonstration of specific markers octamer binding transcription factor and protein gene product.All the morpho-biometrical parameters of left and right testes, except density, were significantly (P<0.01) different between the groups. The total number of SSCs isolated from pre-pubertal testes (6.85×106 cells/ml) was significantly (P<0.05) higher than post-pubertal testes (2.40×106 cells/ml). The colonies of SSCs inpre-pubertal culture appeared within seven days of culture whereas in post-pubertal SSCs, colonies appeared after 14 days of culture. The results demonstrated that the SSCs in prepubertal goat testes were higher in number, grew faster and formed colonies at least one week earlier compared to the SSCs of post-pubertal testes. Thus, the testes of pre-pubertal goats were a better source to study the physiological and regulatory mechanisms of SSCs.