Abstract
ObjectiveMesenchymal stem/stromal cells (MSC) were recently discovered in the human endometrium. These cells possess key stem cell properties and show promising results in small animal models when used for preclinical tissue engineering studies. A small number of surface markers have been identified that enrich for MSC from bone marrow and human endometrium, including the Sushi Domain-containing 2 (SUSD2; W5C5) and CD271 markers. In preparation for developing a large animal preclinical model for urological and gynecological tissue engineering applications we aimed to identify and characterise MSC in ovine endometrium and determine surface markers to enable their prospective isolation.Materials and MethodsOvine endometrium was obtained from hysterectomised ewes following progesterone synchronisation, dissociated into single cell suspensions and tested for MSC surface markers and key stem cell properties. Purified stromal cells were obtained by flow cytometry sorting with CD49f and CD45 to remove epithelial cells and leukocytes respectively, and MSC properties investigated.ResultsThere was a small population CD271+ stromal cells (4.5 ± 2.3%) in the ovine endometrium. Double labelling with CD271 and CD49f showed that the sorted CD271+CD49f- stromal cell population possessed significantly higher cloning efficiency, serial cloning capacity and a qualitative increased ability to differentiate into 4 mesodermal lineages (adipocytic, smooth muscle, chondrocytic and osteoblastic) than CD271-CD49f- cells. Immunolabelling studies identified an adventitial perivascular location for ovine endometrial CD271+ cells.ConclusionThis is the first study to characterise MSC in the ovine endometrium and identify a surface marker profile identifying their location and enabling their prospective isolation. This knowledge will allow future preclinical studies with a large animal model that is well established for pelvic organ prolapse research.
Highlights
Tissue engineering (TE) is the combination of a range of biological and synthetic material scaffolds with a variety of cell types and has revolutionized treatment options for several clinical conditions
Double labelling with CD271 and CD49f showed that the sorted CD271+CD49f- stromal cell population possessed significantly higher cloning efficiency, serial cloning capacity and a qualitative increased ability to differentiate into 4 mesodermal lineages than CD271-CD49f- cells
Since CD271 has been reported on ovine bone marrow MSC (bmMSC) [24], we examined CD49f-CD45ovine endometrial stromal cells for expression of CD271
Summary
Tissue engineering (TE) is the combination of a range of biological and synthetic material scaffolds with a variety of cell types and has revolutionized treatment options for several clinical conditions. TE approaches using stem cells and in particular mesenchymal stem/stromal cells (MSC) are most promising because they possess key properties; self-renewal, high proliferative potential and differentiation. Human endometrium contains a small population of clonogenic stromal cells with typical MSC properties [9,10,11]. Endometrial MSC (eMSC) have been identified as a component of endometrial side-population (SP) cells [11,12,13,14]. The eMSC are clonogenic and self-renew as demonstrated by serial cloning in culture [10]; they undergo multilineage differentiation into four mesenchymal lineages, including smooth muscle cells in vitro, indicating their similarity to bone marrow MSC. One advantage of human eMSC is the relative ease with which they can be obtained by an endometrial biopsy as an office-based procedure without the use of anaesthesia, which is significantly less painful or invasive than bone marrow aspiration or liposuction [16]
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