Optical microscope gross characterization of rat neonatal testicular germline stem cell expansion in vitro

Abstract

Neonatal testicular germline stem cell suspension prepared from twenty 3-5 day old albino rats were incubated in normal saline or in NC-01in triplicate groups. A control group of saline alone (without cell suspension) was also prepared. The cell-containing groups and control were incubated for 21 days at room temperature, stained with 50% Mezo (organic) and visually studied daily with a view to histological characterizing the changes in cellular aggregation, morphology and coloration during incubation. Images of dense oval cells and cellular developmental transformation into mixed populations of these testicular cells showed clearly identifiable patterns of clump formation. Saline alone (control) did not show the presence of any cells during the period of incubation. In vitro expansion of neonatal germline stem cells within 21 days of incubation was associated with multiple divisions into numerous structurally similar cells and transformation into clusters of multi-colored component rod-like cells. A many-fold increase in the number of dense oval cells was observed in the NC-01 culture medium than in the normal saline. Using these morphological criteria and functional characteristics, we suggest a many-fold increased presence of rat neonatal spermatogonial stem cells capable of cellular division in NC-01 culture medium than in normal saline. Key words: Rat, neonatal germline stem cell, Mezo stain, cell suspension, NC-01