<title>Abstract</title> The interplay between stromal and epithelial interactions is pivotal in the development and maintenance of various organs, including the prostate, an essential accessory gland in the male reproductive system. The prostate gland's development and physiology are intricately tied to the localized concentrations of sex steroids, specifically androgens and estrogens. Perturbations in this hormonal balance, often associated with aging, have been implicated in prostatic diseases such as hyperplasia and cancer. In this study, we investigated the influence of estradiol (E2) and dihydrotestosterone (DHT) on prostate cells’ interactions and acinar morphogenesis. Increasing E2:DHT ratios were found to enhance stromal proliferation without a concurrent increase in epithelial cell population. Stromal conditioned medium (CM) promoted aberrant epithelial growth via elevated mitogenic factors, ERα/AR expression, and BAX downregulation. Notable expression of CK05 marker suggests basal cell involvement. Additionally, CM stimulated acinar branching and expansion in prostatospheres, mimicking early cancerous acinar changes. These findings illuminate the complex interplay between hormonal ratios and stromal-epithelial crosstalk, offering valuable insights into potential mechanisms underlying age-related prostate pathologies.