Leucine-rich repeat-containing G-protein-coupled receptor 5-positive cells in the endometrial stem cell niche
Leucine-rich repeat-containing G-protein-coupled receptor 5-positive cells in the endometrial stem cell niche
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DOI：
10.1016/j.fertnstert.2016.10.021
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发表时间：
2017-02-01
影响因子：
6.7
通讯作者：
Simon, Carlos
中科院分区：
文献类型：
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作者：
Cervello, Irene;Gil-Sanchis, Claudia;Simon, Carlos
目的：研究、分离和鉴定人子宫内膜富含亮氨酸重复序列的异三聚体鸟嘌呤核苷酸结合蛋白偶联受体5(LGR5)阳性细胞，以确定其功能相关性。设计：前瞻性实验动物研究。地点：大学科研实验室。动物(S)：非肥胖糖尿病小鼠(NOD-SCID)(品系编码394；NOD。干预(S)：用超顺磁性氧化铁纳米颗粒(SPIO)标记人LGR5(+)细胞，并将其注射到免疫低下小鼠的肾被膜下。主要观察指标(S)：采用荧光激活细胞分选法从人子宫内膜分离LGR5(+)细胞，并使用造血和间质标志物通过流式细胞术进行表型鉴定。植入SPIO标记的LGR5(+)细胞，用普鲁士蓝染色和CD9、Vimentin免疫组织化学方法进行定位。结果：LGR5(+)细胞占子宫内膜上皮和间质细胞总数的比例分别为1.08+/-0.73%和0.82+/-0.76%。LGr5(+)细胞大量表达CD45造血标志，不表达表面标志CD31、CD34、CD133、CD73和CD90。检测到巨噬细胞标志物CD163的共表达。将标记的LGR5(+)细胞异种移植到免疫受损小鼠的肾被膜中，导致该来源细胞的子宫内膜重建较弱。转录谱分析揭示了LGR5(+)细胞与其可能的造血起源相关的新属性。结论：这些数据表明子宫内膜LGR5不是内源性干细胞标记物。相反，LGR5(+)细胞似乎是从血液中招募来的，成为血管周围微环境中干细胞利基的一部分，以激活内源性利基。
Objective: To study, isolate and characterize leucine-rich repeat-containing heterotrimeric guanine nucleotide-binding proteincoupled receptor 5 (LGR5)-positive cells from human endometrium to determine their functional relevance.Design: Prospective experimental animal study.Setting: University research laboratories.Animal(s): Nonobese diabetic mice (NOD-SCID) (strain code 394; NOD. CB17-Prkdcscid/NcrCrl).Intervention(s): Human LGR5(+) cells were labeled with superparamagnetic iron oxide nanoparticles (SPIOs) and injected under the kidney capsule in immunocompromised mice.Main Outcome Measure(s): Epithelial and stromal LGR5(+) cells were isolated from human endometrium by means of fluorescenceactivated cell sorting, and phenotypic characterization was performed by means of flow cytometry with the use of hematopoietic and mesenchymal markers. Engrafted SPIO-labeled LGR5(+) cells were localized with the use of Prussian blue staining and immunohistochemistry against CD9 and Vimentin. Deep transcriptomic profiling of LGR5(+) cells was performed with the use of microarrays and RNA sequencing.Result(s): The percentage of LGR5(+) cells in human endometrium represented 1.08 +/- 0.73% and 0.82 +/- 0.76% of total cells in the epithelial and stromal compartments, respectively. LGR5(+) cells were phenotypically characterized by abundant expression of CD45 hematopoietic marker and no expression of surface markers CD31, CD34, CD133, CD73, and CD90. Coexpression with the macrophage marker CD163 was detected. Xenotransplantation of labeled LGR5(+) cells into the kidney capsules of immunocompromised mice resulted in a weak endometrial reconstitution from this cell of origin. Transcriptomic profiling revealed new attributes for LGR5(+) cells related to their putative hematopoietic origin.Conclusion(s): These data suggest that endometrial LGR5 is not an endogenous stem cell marker. Instead, LGR5(+) cells appear to be recruited from blood to be part of the stem cell niche at the perivascular microenvironment to activate the endogenous niche.