Clonality of smooth muscle and fibroblast cell populations isolated from human fibroid and myometrial tissues
- 15 November 2013
- journal article
- Published by Oxford University Press (OUP) in Molecular Human Reproduction
- Vol. 20 (3), 250-259
- https://doi.org/10.1093/molehr/gat083
Abstract
Uterine fibroids are conventionally defined as clonally derived benign tumours from the proliferation of a single smooth muscle cell (SMC). We have previously identified fibroblast-like cells in fibroids, the presence of which raises the question as to whether all cells within the fibroid have the same clonal origin. The first aim of this study was to develop a fluorescence-activated cell sorting (FACS)-based method to isolate different cell types from human myometrium and fibroid tissues. Secondly, we aimed to use X chromosome inactivation analysis to determine the clonality of cell subpopulations isolated from myometrial and fibroid tissues. Human myometrium and fibroid tissues were collected from women undergoing hysterectomy. Immunohistochemistry (IHC) and flow cytometry confirmed that in addition to SMCs, fibroblasts constitute a significant proportion of cells in human myometrium and fibroid tissues. FACS based on CD90 and ALDH1 reliably separated cells into three myometrial and four fibroid subpopulations: SMCs, vascular smooth muscle cells and two fibroblast subsets. Clonality was first determined by X chromosome inactivation using the classic DNA methylation-sensitive HUMARA assay. Data from this assay were highly variable, with only a quarter of samples meeting the definition of clonal fibroid and non-clonal myometrium. However, using an RNA-based X chromosome inactivation HUMARA assay, we were able to demonstrate clonality of all cellular constituents of most fibroids. Our results confirm that most fibroids are derived from a single cell, and for the first time demonstrates that these clonal cells differentiate into fibroblast and SMC subpopulation as the fibroid grows.Keywords
This publication has 46 references indexed in Scilit:
- Methylation of AR locus does not always reflect X chromosome inactivation stateBlood, 2012
- Evolutionary diversity and developmental regulation of X-chromosome inactivationHuman Genetics, 2011
- Human uterine leiomyoma-derived fibroblasts stimulate uterine leiomyoma cell proliferation and collagen type I production, and activate RTKs and TGF beta receptor signaling in cocultureCell Communication and Signaling, 2010
- Uterine Leiomyomas Exhibit Fewer Stem/Progenitor Cell Characteristics When Compared With Corresponding Normal MyometriumReproductive Sciences, 2009
- CD90 Expression on human primary cells and elimination of contaminating fibroblasts from cell culturesCytotechnology, 2009
- Side population in human uterine myometrium displays phenotypic and functional characteristics of myometrial stem cellsProceedings of the National Academy of Sciences of the United States of America, 2007
- ALDH1 Is a Marker of Normal and Malignant Human Mammary Stem Cells and a Predictor of Poor Clinical OutcomeCell Stem Cell, 2007
- X-linked clonality testing: interpretation and limitationsBlood, 2007
- Fibroblasts in cancerNature Reviews Cancer, 2006
- X-inactivation profile reveals extensive variability in X-linked gene expression in femalesNature, 2005