<![CDATA[The aim of this study was to evaluate the ability of rats Leydig cells collected with Nycodenz gradient in producing testosterone in vitro. Leydig cells were collected using 5 column of Nycodenz gradient (4, 8, 10, 12, and 15%) and cells were evaluated regarding its concentration, viability, and purity of Leydig cells. Media used to cultured Leydig cells were Dulbeccos Modified Eagles Medium (DMEM)+10% newborn calf serum (NBCS); DMEM+10% NBCS+2,5 IU/mL human chorionic gonadotrophin (hCG); DMEM+10% NBCS+ 5 g/mL insulin, 10 g/mL transferrin, and 5 g/mL Se (ITS); DMEM+10% NBCS+hCG+ITS at 5% CO2 incubator with temperature of 37.5 C for 3 days. Culture medium was collected every day for testosterone analysis with enzyme-linked immunosorbent assay (ELISA). By adding ITS to the medium, Leydig cells concentration was significantly increased (8.92x106 cells/mL) compared to medium with serum (7.74x106 cells/mL) or hCG (7.68x106 cells/mL) (P0.05). ITS and hCG in medium significantly increased Leydig cells concentration (10.40x106 cells/mL) at day 3 of culture (P0.05). The result of parallelism test showed that the assay obtained good validity to measure testosterone concentration in culture medium. Testosterone in medium was detected at 1.80-2.60 ng/mL at day 1 of culture. In conclusion, Leydig cells collected with Nycodenz gradient had no effect to testosterone secretion from Leydig cells in vitro.]]>
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