Afami, Marina and Martin, Frank and Fullwood, Nigel (2015) Identify and characterise stem cell population of the human endometrium. Masters thesis, Lancaster University.
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Abstract
The human endometrium is a highly regenerative tissue that experiences functional and structural changes during each menstrual cycle in order to provide a favourable environment for implantation of the embryo. Underpinning this cycle must be a stem cell population. The aim of this project was to characterise this stem cell population in situ in post-menopausal endometrium. To this end, Scanning electron microscopy, image analysis and biospectroscopy techniques, Fourier transform Infrared spectroscopy and Raman spectroscopy, were employed. In a menstruating endometrium, the functional layer is sloughed off during each cycle whilst the basal layer, which contains the potential for endometrial regeneration remains intact. Cyclical and functional changes of the endometrium are regulated by the production of oestrogen and progesterone during the ovarian cycle. Post-menopausal endometrium is thinner and is primarily composed of the basal layer. It is inactive and undergoes apoptotic changes however it retains its regenerative capacity to respond to exogenous hormones. The current evidence supports the hypothesis that epithelial stem/progenitor cells are located in the endometrial crypts, which probably reach the basal layer and migrate to the functional layer to regenerate the epithelial lining of the endometrial glands whilst stromal stem/progenitor cells are located near blood vessels in the functional and basal layer and are responsible for restoring the lost stroma surrounding the endometrial glands. Results from Scanning electron microscopy revealed the different architecture of the endometrial surface in the samples examined. Variations were noticed in cellular morphology as well as in formation of ciliated cells and pinopode-like structures. It was also observed that cells surrounding the endometrial crypts had a more elongated shape relative to the cells away from the crypts which exhibited a more spherical shape. To the best of our knowledge, no prior studies have investigated endometrial stem cell populations by means of biospectroscopy techniques. Spectra were collected from epithelial cells at the base and the lumen of endometrial glands as well as from cells in the surrounding stroma. Pre-processing and subsequent multivariate analysis were applied on the derived spectra to examine segregation between the three populations of cells as well as to characterise their biochemical composition in the hope of identifying spectral regions that discriminate the cell populations and thus support the theory about the putative location of endometrial stem/progenitor cells. Unfortunately, we were not successful in achieving our objectives; however a degree of separation was observed between the different cell types with epithelial cells at the lumen of the glands being the most dissimilar whilst cells at the base of the glands and the surrounding stroma were more similar with respect to each other. Our work though can be used to develop new approaches to further study and provide more insights about stem cell populations in the human endometrium.