163. POTENTIAL MARKERS FOR THE PROSPECTIVE ISOLATION OF HUMAN ENDOMETRIAL EPITHELIAL STEM/PROGENITOR CELLS
C. E. Gargett A B , C. S. C. Tan A B and H. Buhring CA The Ritchie Centre, Monash Institute of Medical Research, Clayton, Australia.
B Obstetrics and Gynaecology, Monash University, Clayton, VIC, Australia.
C CSO Facility for Flow Cytometry and Monoclonal Antibodies, University Clinic of Tubingen, Tubingen, Germany.
Reproduction, Fertility and Development 22(9) 81-81 https://doi.org/10.1071/SRB10Abs163
Published: 6 September 2010
Abstract
The human endometrium regenerates each month following menstruation, parturition and in post-menopausal women taking hormone replacement therapy. Adult stem/progenitor cells discovered residing in human and mouse endometrium may be responsible for this regenerative capacity. However, assays used to identify these stem/progenitor cells are retrospective. The aim of this study is to identify surface markers for the prospective isolation of human endometrial epithelial progenitor cells using a panel of 22 antibodies. Flow cytometry and was used in the initial screen and immunohistochemistry was used to reveal the location of marker expression. Multi-colour FACS protocols were developed with promising markers in conjunction with EpCAM (epithelial cell marker) and to exclude endothelial (CD31+), leukocytes (CD45+) and stromal (CD90+) cells. Sorted subpopulations were assessed for clonogenicity and self-renewal activity using in vitro cloning assays. Six antibodies were short-listed. 2D1D12 enriched for progenitor cells that formed epithelial clones in culture (n = 2). The 2D1D12+EpCAM+ fraction produced very few colony-forming units (CFU). 2D1D12–EpCAM+ fraction gave rise to small CFU. However the 2D1D12+EpCAM– population showed the greatest progenitor activity, producing many large CFU that could be serially cloned twice, indicating self renewal activity. This preliminary data suggests that 2D1D12+EpCAM–CD90–CD31–CD45– population may enrich for human endometrial epithelial stem/progenitor cells. Importantly, large CFU have previously been reported to exhibit stem cell properties of self-renewal, differentiation and high proliferative potential (1). Future studies will focus on xenografting this population to assess tissue reconstitution ability in vivo. The identification of endometrial epithelial stem/progenitor cell marker(s) will enable their prospective isolation for further characterisation and will assist in the investigation of their potential role in endometrial proliferative disorders such as endometriosis and endometrial cancer.
(1) Gargett CE et al (2009) Biol Reprod 80: 1136–45.
