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

Ayako Masuda, Noriko Katoh, Kazuhiko Nakabayashi, Kiyoko Kato, Kenzo Sonoda, Mari Kitade, Satoru Takeda, Kenichiro Hata, Junko Tomikawa

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14 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)213-218
Number of pages6
JournalJournal of Reproduction and Development
Volume62
Issue number2
DOIs
Publication statusPublished - Apr 22 2016

All Science Journal Classification (ASJC) codes

  • Animal Science and Zoology

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    Masuda, A., Katoh, N., Nakabayashi, K., Kato, K., Sonoda, K., Kitade, M., Takeda, S., Hata, K., & Tomikawa, J. (2016). An improved method for isolation of epithelial and stromal cells from the human endometrium. Journal of Reproduction and Development, 62(2), 213-218. https://doi.org/10.1262/jrd.2015-137