Three-dimensional printing of oviductal and endometrial extracellular matrices (ECM) and its implications on in vitro-produced embryos co-culture

• Grant number: 21/11279-2
• Support Opportunities: Scholarships abroad - Research Internship - Master's degree
• Start date: December 28, 2021
• End date: March 27, 2022
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Juliano Coelho da Silveira
• Grantee: Giuliana de Avila Ferronato
• Supervisor: Marcia de Almeida Monteiro Melo Ferraz
• Host Institution: Faculdade de Zootecnia e Engenharia de Alimentos (FZEA). Universidade de São Paulo (USP). Pirassununga , SP, Brazil
• Institution abroad: Ludwig Maximilian University of Munich (LMU Munich), Germany
• Associated to the scholarship: 19/25675-7 - In vitro production of bovine embryos in different 3D culture systems: effects on nuclear morphology, methylation pattern, gene transcription and extracellular vesicles, BP.MS

Abstract

The in vitro embryo production technique (IVEP) was developed almost 40 years ago and still has some limitations that need to be overtaking, such as some diseases that can compromise the animals after birth. During this time the science and technology also had great advances that could be applied to improve IVEP. One of this is 3D bioprinting, a technique that allow the printing of structure that better mimic of in vivo tissues in in in vitro cultures. The ink utilized can be various biological materials, as the extracellular matrix (ECM). ECM is a structurally and stable composite that contributes to the mechanical properties and provides biochemical cues for tissues. So, the objective of this project is to use isolated oviduct and endometrium ECM in bioprinting and co-cultivate it with IVEP embryos. In experiment l, the whole reproductive tract from 5 female cows will be decellularized, and then an endometrial and oviduct fraction tissue will be isolated, milled, and lyophilized. The resulting ECM lyophilized will be digested and neutralized to convert in a hydrogel. For 3D printing, we will create a gelatin slurry support bath were the ECM hydrogel could be impress without melt its structure. We will use the BIO X bioprinter (CELLINK, Sweden) and ECM hydrogel as bioink. The 3D printing will have the oviduct or endometrium folds format. After that, this bioprint will be characterized to proceed with experiment ll. In experiment II, Cumulus Oocyte Complexes (COC) from local slaughterhouse ovaries will be matured and fertilized, on the day of cultivation they will be divided into 3 groups: 1) conventionally 2D IVEP as a control; 2) IVEP co-culture with 3d bioprinted ovidutal ECM; and 3) IVEP co-culture with 3D bioprinted endometrial ECM. In the student's Master project in Brazil, there is also the intention to search for better and new models for IVEP and this is how the two projects are related and can be compared. So, we expect from these results to find a new model for IVEP that can improve embryo quality and overcame the actual needs of the technique.