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 language | English |
|---|---|
| Pages (from-to) | 213-218 |
| Number of pages | 6 |
| Journal | Journal of Reproduction and Development |
| Volume | 62 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - Apr 22 2016 |
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All Science Journal Classification (ASJC) codes
- Animal Science and Zoology
Cite this
An improved method for isolation of epithelial and stromal cells from the human endometrium. / Masuda, Ayako; Katoh, Noriko; Nakabayashi, Kazuhiko; Kato, Kiyoko; Sonoda, Kenzo; Kitade, Mari; Takeda, Satoru; Hata, Kenichiro; Tomikawa, Junko.
In: Journal of Reproduction and Development, Vol. 62, No. 2, 22.04.2016, p. 213-218.Research output: Contribution to journal › Article
}
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
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.
UR - http://www.scopus.com/inward/record.url?scp=84964303637&partnerID=8YFLogxK
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U2 - 10.1262/jrd.2015-137
DO - 10.1262/jrd.2015-137
M3 - Article
VL - 62
SP - 213
EP - 218
JO - Journal of Reproduction and Development
JF - Journal of Reproduction and Development
SN - 0916-8818
IS - 2
ER -