The Ritchie Centre, Monash Institute of Medical Research, and Department of Obstetrics and Gynaecology, Monash University, Clayton, Victoria, Australia.
Cell Transplant. 2012;21(10):2201-14. doi: 10.3727/096368911X637362. Epub 2012 Mar 27.
Coexpression of CD140b (PDGFRβ) and CD146 has been used to isolate endometrial mesenchymal stem-like cells (eMSCs), which have a perivascular location. This study aims to evaluate a single marker for purifying eMSCs. Using an antibody panel with novel specificities, we screened human endometrial tissues and stromal cell suspensions by flow cytometry and immunohistochemistry to identify perivascular markers. Sorted subpopulations were examined for colony-forming unit (CFU), self-renewal, and differentiation assays for mesenchymal stem cell (MSC) function. We also transplanted sorted eMSCs under the kidney capsule of superimmunodeficient NSG mice. Magnetic bead selection was compared with flow cytometry sorting (flow sorting) using CFU assay. One novel marker (W5C5) was particularly effective in selecting eMSCs. W5C5(+) cells comprise 4.2±0.6% (n = 34) of endometrial stromal cells and reside predominantly in a perivascular location in both basal and functional layers of endometrium. The clonogenicity of W5C5(+) cells is significantly greater than W5C5(-) and unselected cells. W5C5(+) cells differentiated into adipocytes, osteocytes, chondrocytes, myocytes, and endothelial cells. W5C5(+) cells produce endometrial stromal-like tissue in vivo. In terms of clonogenicity, magnetic bead-selected W5C5(+) cells gave rise to significantly higher CFU numbers compared to flow-sorted W5C5(+) cells. This study identified W5C5 as a single marker capable of purifying eMSCs possessing MSC properties and reconstituting endometrial stromal tissues in vivo. W5C5 enriches eMSCs to high purity and provides a simple protocol for their prospective isolation using magnetic bead selection rather than flow sorting. W5C5 selection may provide an alternate, readily available autologous source of MSC, obtainable with minimal morbidity using an office endometrial biopsy procedure for future cell-based therapies.
CD140b（血小板衍生生长因子受体β）和 CD146 的共表达已被用于分离具有血管周位置的子宫内膜间充质干细胞（eMSCs）。本研究旨在评估一种用于纯化 eMSCs 的单一标志物。我们使用具有新型特异性的抗体面板，通过流式细胞术和免疫组织化学筛选人子宫内膜组织和基质细胞悬浮液，以鉴定血管周标志物。分选的亚群进行集落形成单位（CFU）、自我更新和间充质干细胞（MSC）功能分化测定。我们还将分选的 eMSCs 移植到超免疫缺陷 NSG 小鼠的肾包膜下。磁珠分选与 CFU 测定的流式细胞分选（流式分选）进行了比较。一种新型标志物（W5C5）在选择 eMSCs 方面特别有效。W5C5(+)细胞占子宫内膜基质细胞的 4.2±0.6%（n = 34），主要位于子宫内膜的基础层和功能层的血管周位置。W5C5(+)细胞的克隆形成能力明显大于 W5C5(-)和未分选细胞。W5C5(+)细胞可分化为脂肪细胞、成骨细胞、软骨细胞、肌细胞和内皮细胞。W5C5(+)细胞在体内产生子宫内膜基质样组织。在克隆形成能力方面，与流式分选的 W5C5(+)细胞相比，磁珠分选的 W5C5(+)细胞产生的 CFU 数量显著更高。本研究鉴定出 W5C5 是一种能够纯化具有 MSC 特性并在体内重建子宫内膜基质组织的 eMSCs 的单一标志物。W5C5 可将 eMSCs 高度纯化，并提供一种简单的方案，通过磁珠选择而不是流式分选来进行其前瞻性分离。W5C5 选择可能提供一种替代的、易于获得的 MSC 自体来源，通过对办公室子宫内膜活检程序进行最小的发病率操作，可获得用于未来细胞治疗的 MSC。
