A Novel Marker of Human Endometrial Mesenchymal Stem-Like Cells

Abstract

Coexpression of CD140b (PDGFRb) 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. + 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.

Angiogenic properties of endometrial mesenchymal stromal cells in endothelial co-culture: an in vitro model of endometriosis

“…In addition, clonally purified SUSD2 + E-MSCs were shown to acquire 88 endothelial marker expression and to integrate into renal blood vessels after xenograft under the 89 kidney capsule, underlying an endothelial differentiative ability (Masuda et al, 2012 for 48 hours. HUVECs and E-E-s were also cultured alone and used as control.…”

Regeneration capacity of the endometrium: endometrial stem cells

“…[9] Endometrium içerisin-de bazalis ve fonksiyonalis tabakaları içerisindeki damarlar çevresinde yerlemi olan kemik ili¤i kaynaklı mezenkimal kök hücreler ile benzer yüzey belirteçleri gösteren ve aynı zamanda PDGFR-b (platelet derived growth factor receptor-b) ve SUSD2 (sushi domain containing 2) gibi belirteçle-re de sahip olan hücreler endometrial mezenkimal kök hücreler olarak tanımlanmılardır ve bu hücre-lerin kemik ili¤i kaynaklı mezenkimal kök hücreler ile in vitro koullarda benzer özellikler gösterdik-leri saptanmıtır. [12][13][14] Yapılan bir ksenogreft model çalımasında SUSD2+ endometrial mezenkimal kök hücrelerin immün sistemi baskılanmı farelerin böbrek kapsülü altına yerletirildiklerinde, renal parankimal damar duvarları çevresinde endometial stroma oluturabildikleri gözlenmitir. [10] Yapılan çalımalarda endometrial stroma içeri-sinde CD146+, PDGFR-b+ hücrelerin endometrial stromal hücrelerinin %1.5'ini oluturdu¤u gösterilmitir.…”

“…Yapılan çalımalarda endometrial stromal LRC hücrelerinin endometrial-myometrial bilekede luminal epitelyumun altında veya perivasküler bölgede yerletikleri gösterilmitir. [12] Bu hücrelerin kök hücre belirteçlerinden Oct-4, c-Kit, [13] CD 140, CD146, CD44, CD90 ve Sall4'ü eksprese ettikleri saptanmıtır. [15,16] Epiteliyal LRC hücreleri, epitelial hücrelerin yüksek ço¤alma öze-liklerinden dolayı çok kısa bir süre persiste ettiklerinden zor tespit edilmektedirler.…”

Isolation and Primary Culture of Various Cell Types from Whole Human Endometrial Biopsies

“…They maintain many mesenchymal stem cell characteristics (Masuda et al, 2012), and can be cultured and differentiated (Ulrich et al, 2014) and assessed for clonogenicity using CFU (Colony Forming Units) assay (Masuda et al, 2012;Murakami et al, 2013; as required. Figure 2C).…”

“…Furthermore, within dissociated HESCs there is a sub-population of perivascular progenitor mesenchymal stem-like cells that can be isolated using the perivascular-specific antigen SUSD2 and its cognate antibody W5C5. Here we provide in detail an updated and expanded protocol from those published previously (Masuda et al, 2012;Chen and Roan, 2015) to describe steps in isolating and culturing different cell types from whole human endometrium. We provide further information on biopsy collection, detailed protocols for isolation of progenitor cells and additional procedures to increase epithelial cell yield and culturing efficiency.…”