The main functions of the testis are sex hormone and sperm cell production. Steroidogenesis occurs in the Leydig interstitial cells and spermatogenesis in the seminiferous tubules. Male gonad morphogenesis is a finely orchestrated process, mainly coordinated by hormones, whose actions can significantly affect post‐pubertal testicular function. Calcium is a key intracellular messenger, which regulates many signal transduction pathways, and is also implicated in steroidogenesis. Calcium homeostasis and signaling rely on many calcium‐binding proteins including calretinin, of the “EF‐hand” protein family. Calretinin is a highly conserved protein mainly expressed in the nervous system but also detected in rat and human adult and fetal testis as well as in pathological conditions. Calretinin expression in the fetal testis, however, has not been thoroughly analyzed probably owing to limited availability and paucity of tissues. Here, we examined by immunocytochemistry the expression of calretinin in human fetal testis specimens, obtained from natural and therapeutic abortions, at various developmental ages. We found that calretinin‐immunoreactive Leydig cells were visible throughout the timeframe studied (14th–27th week). Immunoreactivity was also observed in Sertoli cells and in the germ cells of the immature seminiferous tubules. Overall our data indicate that calretinin expression parallels the decline in Leydig cell number, suggesting that its presence is indeed correlated to their steroidogenic activity. They also suggest that the intratubular positivity of calretinin could be linked to the ability of Sertoli cells to produce locally acting hormones contributing to the histodifferentiation of the male genital tract. J. Cell. Physiol. 232: 1872–1878, 2017. © 2016 Wiley Periodicals, Inc. Human fetal testis specimens, obtained upon natural and therapeutic abortions at various developmental ages (weeks 14–27), show calretinin expression in the Leydig interstitial cells, in the epididymis and in the immature seminiferous tubules. Calretinin expression parallels the decline in Leydig cell number. The presence of calretinin seems correlated to the Leydig cells steroidogenic activity, which is consistent with the role that calcium intracellular signaling plays in the steroidogenic process. Our findings, by adding calretinin to the list of players underlying fetal testis development, are particularly timely and prompt further assessment of calretinin role in this context both in normal and pathological conditions.