Rawlings, Thomas (2020) Three-dimensional modelling of the human peri-implantation endometrium. PhD thesis, University of Warwick.

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Abstract

Human embryo implantation is the rate-limiting step for successful pregnancy in assisted reproductive technology, and inappropriate endometrial embryo-quality biosensing is also associated with recurrent pregnancy loss. Elucidating the mechanisms underpinning the peri-implantation endometrium would enable the development of treatments for patients suffering with chronic fertility disorders. However, implantation is difficult to study. Therefore, simple in vitro models of human endometrial cells have been developed. However, the endometrium is a complex tissue made up of stromal cells (EnSC), epithelial glands, vasculature, and immune cells. An in vitro model that encompasses all aspects of the human endometrium would require a complex co-culture system. Therefore, three-dimensional (3D) culturing techniques are the most suitable. This study aimed to establish a protocol for a novel in vitro model of the peri-implantation endometrium by co-culturing primary EnSC and epithelial cells (EEC) using 3D culture techniques. In this study, two approaches were undertaken. The first involved the use of the Endometrial Regenerative Bodies (ERB) model, which was unsuccessful in achieving the study’s aim. However, the second approach, involving the 3D co-culture of EnSC with endometrial gland organoids led to the establishment of a complex organoid culture. The growth conditions of the complex organoids were optimised by modifying the 3D matrix and medium conditions. The standard organoid growth matrix, Matrigel, a basement membrane derived hydrogel, was replaced with a purified Type I collagen hydrogel to provide a more physiological ECM for the EnSC. Direct interaction between the EnSC and EEC in the complex organoids was examined through the establishment of a minimal differentiation medium and single cell RNA-sequencing of a decidual time course of gland organoids and complex organoids. EnSC were able to induce the differentiation of EEC into a secretory phenotype in complex organoid cultures. The established complex organoid protocol offers a starting point for developing a faithful in vitro model of the human peri-implantation endometrium, which will provide a step change in our collective ability to study human embryo implantation. A system that would be patient-specific, and in the long term developed into a high-throughput system for personalised medicine for the treatment of infertility.

Item Type:	Thesis (PhD)
Subjects:	R Medicine > RG Gynecology and obstetrics
Library of Congress Subject Headings (LCSH):	Human embryo -- Transplantation, Endometrium, Cell culture, Cytology -- Technique, Fertilization in vitro, Human
Official Date:	January 2020
Dates:	Date Event January 2020 UNSPECIFIED
Institution:	University of Warwick
Theses Department:	Warwick Medical School
Thesis Type:	PhD
Publication Status:	Unpublished
Supervisor(s)/Advisor:	Brosens, Jan J.
Format of File:	pdf
Extent:	xiii, 245 leaves : illustrations, charts
Language:	eng

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