Use of extracellular matrix of bovine endometrium for hydrogel formation as an in vivo embryonic maternal communication model

Abstract

Much has been discussed about embryonic loss in dairy and beef cattle, such losses are directly related to the failure in maternal embryonic communication in the uterus, and better understanding this communication is key to improve in vitro embryo production. Three-dimensional culture of endometrial cells, in which cells form a three-dimensional structure, improving intercellular communication and acting more physiologically, have been used to understand this environment. ECM is a key point for this type of culture, as it presents a structurally and stable composition that contributes to the mechanical properties and provides biochemical cues for tissues. So, the main objective of this project is to isolate endometrial ECM to generate an ECM hydrogel (EndoECM) to establish organoid cultures from endometrial glands for an in vitro study model of embryonic maternal communication. In experiment l, we will decellularize uterus pieces, and then endometrial fraction tissue will be isolated, milled, and lyophilized. The resulting lyophilized ECM will be digested and neutralized to convert into a hydrogel and will be characterized. After the characterization, we will proceed to experiment II. In experiment II, we will carry out the culture of glandular epithelial cells for the formation of organoids that will be co-cultured with epithelial and stromal cells. After their formation and co-culture, we will start the last experiment. Experiment III will be conducted using the co-cultured organoids form Experiment II to generate a maternal environment for in vitro embryo production (IVEP). The embryos will be produced in vitro under two conditions: 1) IVEP alone; 2) IVEP co-cultured with endoECM and surrounded by an epithelial and stromal cell monolayer. The analyses that will be performed in this project are the same ones used in the student's FAPESP scholarship in Brazil. As a result, the data obtained can be compared to that of other groups in order to determine the best model for use in the communication between maternal and embryonic tissues.