TY - JOUR
T1 - An improved method for isolation of epithelial and stromal cells from the human endometrium
AU - Masuda, Ayako
AU - Katoh, Noriko
AU - Nakabayashi, Kazuhiko
AU - Kato, Kiyoko
AU - Sonoda, Kenzo
AU - Kitade, Mari
AU - Takeda, Satoru
AU - Hata, Kenichiro
AU - Tomikawa, Junko
N1 - Publisher Copyright:
© 2016 by the Society for Reproduction and Development.
PY - 2016/4/22
Y1 - 2016/4/22
N2 - 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 fluorescenceactivated 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.
AB - 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 fluorescenceactivated 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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U2 - 10.1262/jrd.2015-137
DO - 10.1262/jrd.2015-137
M3 - Article
C2 - 26853786
AN - SCOPUS:84964303637
VL - 62
SP - 213
EP - 218
JO - Journal of Reproduction and Development
JF - Journal of Reproduction and Development
SN - 0916-8818
IS - 2
ER -