Gene expression profiling in human endometrial cells during hormonal stimulation

Wiid, Kyle
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The menstrual cycle is a complex system of dynamically interacting hormones, growth factors, cytokines and cells. The cycle is regulated by the combined effects of both the 17β-oestradiol (E2) and progesterone hormones. An in-depth understanding of complex systems can be obtained if all of the processes that constitute the system are evaluated individually. Therefore, this study aimed to assess the individual effects of both E2 and medroxyprogesterone acetate (MPA) on human endometrial stromal cells (HESCs) in vitro. The effects of these hormones on the expression of ESR-α, PGR, TGF-β1 and VEGF A was assessed using Real-Time qRT-PCR, and the secretion of VEGF A and IL-6 was assessed using ELISA. A successful and financially feasible methodology for the isolation of HESCs using endometrial biopsies was established. The cryopreservation technique used in this study resulted in low cell viability, and the data could not be statistically validated. However, the results do show similarities to published data. In the presence of E2, ESR-α expression was minimally altered, and PGR expression was minimally increased. Additionally, an increase in growth factor expression was observed. VEGF A was upregulated in the presence of both E2 and MPA, whereas TGF-β1 was only upregulated in the presence of E2. Despite the fact that the data could not be statistically validated, the study provides valuable insight into the individual actions of E2 and MPA on the menstrual cycle. Furthermore, the study highlights the importance of cryopreservation in the discipline of cell culture
A Dissertation submitted to the Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, in fulfilment of the requirements for the degree of Master of Science by Dissertation October , 2019 Johannesburg