一种从人子宫内膜分离上皮细胞和基质细胞的改进方法

Ayako Masuda, Noriko Katoh, K. Nakabayashi, Kiyoko Kato, K. Sonoda, Mari Kitade, S. Takeda, K. Hata, Junko Tomikawa
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引用次数: 23

摘要

我们旨在提高从人子宫内膜中分离子宫内膜上皮细胞和基质细胞(EMECs和EMSCs)的效率。我们通过免疫组织化学染色发现,胶原酶处理后遗留的大块组织碎片由腺上皮细胞和基质细胞组成,这些组织碎片通常在常规方案中第一次过滤后被丢弃。因此,我们建立了蛋白酶处理和细胞悬浮条件,通过荧光激活细胞分选将单个细胞从组织片段和分离的上皮细胞(epcam阳性)和基质细胞(cd13阳性)中分离出来。4项独立实验结果显示,1个子宫切除标本平均分离EMECs 1.2 × 106个,EMSCs 2.8 × 106个。我们证实,分离的细胞表现出与常规方法获得的上皮细胞和基质细胞高度相似的转录组特征。我们改进的方案有助于未来的研究,以更好地了解子宫内膜在月经周期中动态变化的分子机制。
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An improved method for isolation of epithelial and stromal cells from the human endometrium
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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