An improved method for isolation of epithelial and stromal cells from the human endometrium

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  • MASUDA Ayako
    Department of Maternal-Fetal Biology, National Research Institute for Child Health and Development, Tokyo 157-8535, Japan Department of Obstetrics and Gynecology, Faculty of Medicine, Juntendo University, Tokyo 113-8431, Japan
  • KATOH Noriko
    Department of Maternal-Fetal Biology, National Research Institute for Child Health and Development, Tokyo 157-8535, Japan Department of Obstetrics and Gynecology, Faculty of Medicine, Juntendo University, Tokyo 113-8431, Japan
  • NAKABAYASHI Kazuhiko
    Department of Maternal-Fetal Biology, National Research Institute for Child Health and Development, Tokyo 157-8535, Japan
  • KATO Kiyoko
    Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-0054, Japan
  • SONODA Kenzo
    Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-0054, Japan
  • KITADE Mari
    Department of Obstetrics and Gynecology, Faculty of Medicine, Juntendo University, Tokyo 113-8431, Japan
  • TAKEDA Satoru
    Department of Obstetrics and Gynecology, Faculty of Medicine, Juntendo University, Tokyo 113-8431, Japan
  • HATA Kenichiro
    Department of Maternal-Fetal Biology, National Research Institute for Child Health and Development, Tokyo 157-8535, Japan
  • TOMIKAWA Junko
    Department of Maternal-Fetal Biology, National Research Institute for Child Health and Development, Tokyo 157-8535, Japan

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説明

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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