Trophoblast-endothelial cells co-culture in a 3D system mimicking the embryo implantation to study the trophoblast's role in inducing apoptosis

• Grant number: 20/14982-3
• Support Opportunities: Scholarships in Brazil - Master
• Start date: April 01, 2021
• End date: May 31, 2023
• Field of knowledge: Biological Sciences - Morphology - Histology
• Principal Investigator: Estela Maris Andrade Forell Bevilacqua
• Grantee: Bárbara Stefany da Silva Souza
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Introduction: During the initial stages of implantation in humans, the syncytiotrophoblast and the cytotrophoblast cells interact with uterine endothelial cells. This interaction is believed to include both the establishment of junctions between syncytiotrophoblast membranes and capillary endothelial cells, as well as the induction of endothelial cell death for the remodeling of uterine arterial vessels and the establishment of low-resistance and high-flow channels. These processes occur in a restricted and coordinated way by the trophoblastic populations. Objectives: This study aims to characterize a 3D study model for the analysis of maternal-embryonic interactions, represented here by the interface: trophoblastic cells - co-cultured human endometrial endothelial cells. In addition, this model will be used to evaluate the role of the trophoblast in inducing apoptosis pathways in endothelial cells. Material and methods: For the execution of the project, twenty blastocysts will be provided by the Huntington Clinic of Reproductive Medicine following pre-established inclusion standards. The glandular endometrial epithelium will be isolated from female biopsies of couples with male infertility factor and with indication of treatment by in vitro fertilization, collected according the ethics research protocols (ICB-USP, CONEP). The purity of the uterine samples will be evaluated by immunofluorescence for the following proteins: vimentin, cytokeratin, IGFBP1 and Von Willebrand factor. The 3D coculture system will include the preparation of an extracellular matrix composed of collagen, fibronectin and hyaluronic acid in a Transwell system, on which the isolated uterine endothelial cells will be plated. After stabilization and characterization of this culture, the system will receive blastocysts (n = 2 / assay) on the top of endothelial cells layer (n = 10 experiments). The evaluation of the coculture will be carried out by ultrastructural and immunohistochemical morphological analysis. To detect cells in apoptosis, the presence of cleaved caspase-3, will be evaluated by immunofluorescence using confocal microscopy. (AU)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

SOUZA, Bárbara Stefany da Silva. Characterization of human endometrial cells in a 3D system: the model and its study possibilities. 2024. Master's Dissertation - Universidade de São Paulo (USP). Instituto de Ciências Biomédicas (ICB/SDI) São Paulo.