A refined method for high-purity isolation of uterine glandular epithelial cells in mouse

The uterine endometrium consists of luminal epithelium, glandular epithelium, and stromal cells, with uterine glands playing a pivotal role in pregnancy success among mammals. Uterine glands secrete essential factors that regulate embryo development and implantation; however, their cellular biology remains poorly understood. This study presents a refined method for isolating three distinct endometrial cell types with high purity, with a specific emphasis on glandular epithelial cells. The method combines mechanical dissociation, enzymatic digestion, and immunomagnetic separation. The isolated glandular epithelial cells were maintained in culture and exhibited proliferation in response to steroid hormones. Furthermore, estrogen responsiveness was abrogated by Estrogen Receptor 1 (Esr1) knockdown mediated by siRNA. Here, we present an efficient and reproducible method for isolating uterine glandular epithelial cells with high purity, enabling their in vitro maintenance, hormone responsiveness assessment, and functional gene knockdown. These findings establish a robust platform for advancing our understanding of uterine gland biology, facilitating detailed investigations into molecular mechanisms underlying glandular function and their critical roles in establishing pregnancy success. Future research could explore the contribution of these isolated cells to endometrial receptivity and embryo implantation.