title: Isolation and characterisation of mesenchymal stem/stromal cells in the ovine endometrium.
abstract: OBJECTIVE
  Mesenchymal 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.
  
  
  METHODS
  Ovine 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.
  
  
  RESULTS
  There 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.
  
  
  CONCLUSIONS
  This 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.
authors.affiliation: The Ritchie Centre, MIMR-PHI Institute of Medical Research, Clayton, Victoria Australia.
authors.forename: Vincent
authors.initials: V
authors.lastname: Letouzey
authors.affiliation: The Ritchie Centre, MIMR-PHI Institute of Medical Research, Clayton, Victoria Australia.
authors.forename: Ker Sin
authors.initials: KS
authors.lastname: Tan
authors.affiliation: The Ritchie Centre, MIMR-PHI Institute of Medical Research, Clayton, Victoria Australia.
authors.forename: James A
authors.initials: JA
authors.lastname: Deane
authors.affiliation: The Ritchie Centre, MIMR-PHI Institute of Medical Research, Clayton, Victoria Australia; Department of Obstetrics and Gynaecology, Monash University, Clayton, Victoria Australia.
authors.forename: Daniela
authors.initials: D
authors.lastname: Ulrich
authors.affiliation: The Ritchie Centre, MIMR-PHI Institute of Medical Research, Clayton, Victoria Australia.
authors.forename: Shanti
authors.initials: S
authors.lastname: Gurung
authors.affiliation: The Ritchie Centre, MIMR-PHI Institute of Medical Research, Clayton, Victoria Australia.
authors.forename: Y Rue
authors.initials: YR
authors.lastname: Ong
authors.affiliation: The Ritchie Centre, MIMR-PHI Institute of Medical Research, Clayton, Victoria Australia; Department of Obstetrics and Gynaecology, Monash University, Clayton, Victoria Australia.
authors.forename: Caroline E
authors.initials: CE
authors.lastname: Gargett
mesh_terms: D000068816:Adapalene / Q000378:metabolism*; D000818:Animals; D015415:Biomarkers / Q000378:metabolism*; D002417:Cattle; D002454:Cell Differentiation; D004717:Endometrium / Q000166:cytology*; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D023822:Tissue Engineering
chemical_list: D015415:Biomarkers; D000068816:Adapalene