| Summary: | We aimed to improve the efficiency of isolating endometrial epithelial and stromal cells (EMECs and EMSCs)
from the human endometrium. We revealed by immunohistochemical staining that the large tissue fragments
remaining after collagenase treatment, which are usually discarded after the first filtration in the
conventional protocol, consisted of glandular epithelial and stromal cells. Therefore, we established protease
treatment and cell suspension conditions to dissociate single cells from the tissue fragments and isolated
epithelial (EPCAM-positive) and stromal (CD13-positive) cells by fluorescence-activated cell sorting. Four
independent experiments showed that, on average, 1.2 × 106 of EMECs and 2.8 × 106 EMSCs
were isolated from one hysterectomy specimen. We confirmed that the isolated cells presented transcriptomic
features highly similar to those of epithelial and stromal cells obtained by the conventional method. Our
improved protocol facilitates future studies to better understand the molecular mechanisms underlying the
dynamic changes of the endometrium during the menstrual cycle.
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