More than ever before, people around the world are frequently exposed to different sections of the electromagnetic spectrum, mainly emitted from wireless modern communication technologies. Especially, the level of knowledge on non-thermal biological EMF effects remains controversial. New technologies allow for a more detailed detection of non-coding RNAs which affect the post-transcriptional control. Such method shall be applied in this work to investigate the response of human blood cells to electromagnetic irradiation. In this ex vivo in vitro study, we exposed peripheral blood cells from 5 male donors to a continuous wave of 900 MHz EMF for 0, 30, 60 and 90 min. Significant micro RNA (miRNA) expression changes (p ≤ 0.05) above or below the SHAM exposed samples were evaluated using a quantitative real time PCR platform for simultaneous detection of 667 miRNAs called low density array. Only significant miRNA expression changes which were detectable in at least 60% of the samples per exposure group were analyzed. The results were compared with data from room temperature + 2 °C (RT + 2 °C) samples (here referred to as hyperthermia) to exclude miRNA expression altered by hyperthermia. The validation study by using the same donors and study design was performed after an interval of 2 years. When analyzing a total of 667 miRNAs during the screening study, 2 promising candidate miRNAs were identified, which were down regulated almost twice and showed a complete separation from the unexposed control group (miR-194 at 30 min and miR-939 at 60 min). The p-values even survived the Bonferroni correction for multiple comparisons (p = 0.0007 and p = 0.004, respectively). None of these miRNAs were expressed at a second time point after EMF exposure. Following an alternative analysis approach, we examined for miRNAs revealing an expected significant association of differential miRNA expression with the dose-time EMF exposure product, separately for each donor. Donors 2 and 3 revealed 11 and 10 miRNA species being significantly associated with EMF exposure which differed significantly from the other donors showing a minor number of differentially expressed miRNAs and could identify donors 2 and 3 as particularly EMF-responsive. The measurements were repeated after 2 years. The number of expressed/non-expressed miRNAs was almost similar (97.4%), but neither the number nor the previously differentially expressed miRNAs could be reproduced. Our data neither support evidence of early changes at miRNA expression level in human whole blood cells after 900 MHz EMF exposure nor the identification of EMF-responsive individuals.