An improved method for isolation of epithelial and stromal cells from the human endometrium
| Autor: | Mari Kitade, Junko Tomikawa, Noriko Katoh, Kenzo Sonoda, Kiyoko Kato, Ayako Masuda, Kazuhiko Nakabayashi, Kenichiro Hata, Satoru Takeda |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Adult Stromal cell Cell separation Cellular differentiation Cell Culture Techniques Biology CD13 Antigens Endometrial stromal cells (EMSCs) Endometrium Flow cytometry 03 medical and health sciences chemistry.chemical_compound Young Adult medicine Humans Technology Report Menstrual Cycle medicine.diagnostic_test Gene Expression Profiling Epithelial cell adhesion molecule Cell Differentiation Epithelial Cells Cell sorting Middle Aged Epithelial Cell Adhesion Molecule Flow Cytometry Molecular biology Immunohistochemistry 030104 developmental biology medicine.anatomical_structure chemistry Cell culture Immunology Animal Science and Zoology Female Stromal Cells Transcriptome Endometrial epithelial cells (EMECs) Gene expression array |
| Zdroj: | The Journal of Reproduction and Development |
| ISSN: | 1348-4400 |
| Popis: | 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 × 10(6) of EMECs and 2.8 × 10(6) 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. |
| Databáze: | OpenAIRE |
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