Figure 1.

The influence of various culture media on the capacity of the Leydig cells in three-dimensional cultures of rat testicular cells to produce testosterone and express steroidogenic genes. (A) On the X-axis are the different culture media employed [DMEM + glutamine (GL), F12, F12 + NEAA (non-essential amino acids), F12 + AA (essential amino acids), F12/DMEM, and MEM (minimal essential medium)], and the Y-axis depicts the concentration of testosterone (evaluated by radioimmunoassay and expressed in nanomoles/liter) in the medium of cells cultured for 1 day. The relative expression of (B) Star (Steroidogenic Acute Regulatory Protein) and (C) Tspo (Translocator Protein) (determined by qPCR analysis with Actb as an internal control) by testicular cell suspensions from 7 dpp rats cultured for 1 day in six different media in the presence (light columns) or absence (dark columns) of hCG and FSH. The mean relative expression for triplicates was calculated by the ddCt procedure. One-way ANOVA with the Shapio–Wilk test for normality was applied to compare the different experimental conditions. NS: non-significant; *p < 0.05, **p < 0.01, ***p < 0.001 in comparison to the value with DMEM + glutamine.