Spermatogonial Stem Cells (SSCs) from pre and post-pubertal age groups of mice were isolated by two step enzymatic digestion method and cultured in six-well plates. The in-vitro culture morphology of the SSCs was observed up to passage 2. In primary culture (P0), on day two, the cells were single and transparent. On day 4, the colony forming unit consisted of four to five cells. Three dimensional colonies were observed on day seven post incubation. From day eight onwards, the cells had spindle shape and showed varied morphology till twelve days post incubation. Double nucleolus is prominent in cells on day eight post incubation. In passage 1, though the cells exhibited different morphology viz., round, stellate, polyhedral, and triangular, the stellate cells were found to be more. In passage 2, the spindle-shaped cells with prominent nucleoli were often observed. Other cell morphology like round, stellate, polyhedral and triangular cells were also observed.