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1 June 2009 Isolation and Culture of Epithelial Progenitors and Mesenchymal Stem Cells from Human Endometrium
Caroline E. Gargett, Kjiana E. Schwab, Rachel M. Zillwood, Hong P. T. Nguyen, Di Wu
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Abstract

Human endometrium is a highly regenerative tissue undergoing more than 400 cycles of growth, differentiation, and shedding during a woman's reproductive years. Endometrial regeneration is likely mediated by adult stem/progenitor cells. This study investigated key stem cell properties of individual clonogenic epithelial and stromal cells obtained from human endometrium. Single-cell suspensions of endometrial epithelial or stromal cells were obtained from hysterectomy tissues from 15 women experiencing normal menstrual cycles, and were cultured at clonal density (10 cells/cm2) or limiting dilution. The adult stem cell properties—self-renewal, high proliferative potential, and differentiation of single epithelial and stromal cells—were assessed by harvesting individual colonies and undertaking serial clonal culture, serial passaging, and culture in differentiation-induction media, respectively. Lineage differentiation markers were examined by RT-PCR, immunocytochemistry, and flow cytometry. Rare single human endometrial EpCAM epithelial cells and EpCAM stromal cells demonstrated self-renewal by serially cloning >3 times and underwent >30 population doublings over 4 mo in culture. Clonally derived epithelial cells differentiated into cytokeratin gland-like structures in three dimensional culture. Single stromal cells were multipotent, as their progeny differentiated into smooth muscle cells, adipocytes, chondrocytes, and osteoblasts. Stromal clones expressed mesenchymal stem cell (MSC) markers ITGB1 (CD29), CD44, NT5E (CD73), THY1 (CD90), ENG (CD105), PDGFRB (CD140B), MCAM (CD146) but not endothelial or hemopoietic markers PECAM1 (CD31), CD34, PTPRC (CD45). Adult human endometrium contains rare epithelial progenitors and MSCs, likely responsible for its immense regenerative capacity, which may also have critical roles in the development of endometriosis and endometrial cancer. Human endometrium may provide a readily available source of MSCs for cell-based therapies.

Caroline E. Gargett, Kjiana E. Schwab, Rachel M. Zillwood, Hong P. T. Nguyen, and Di Wu "Isolation and Culture of Epithelial Progenitors and Mesenchymal Stem Cells from Human Endometrium," Biology of Reproduction 80(6), 1136-1145, (1 June 2009). https://doi.org/10.1095/biolreprod.108.075226
Received: 2 December 2008; Accepted: 1 February 2009; Published: 1 June 2009
JOURNAL ARTICLE
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KEYWORDS
Adult stem cells
clonal assays
differentiation
endometrial stem cell
epithelial progenitor cell
female reproductive tract
human endometrium
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