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Simultaneous biopsy and lipid profile of embryo inner cell mass and trofectoderm

Grant number: 19/17796-9
Support Opportunities:Scholarships in Brazil - Doctorate
Effective date (Start): June 01, 2021
Effective date (End): September 30, 2023
Field of knowledge:Agronomical Sciences - Veterinary Medicine - Animal Reproduction
Principal Investigator:Mateus José Sudano
Grantee:Thamiris Vieira Marsico
Host Institution: Centro de Ciências Naturais e Humanas (CCNH). Universidade Federal do ABC (UFABC). Ministério da Educação (Brasil). Santo André , SP, Brazil
Associated research grant:18/21496-8 - Pathways for embryo-derived pregnancy, AP.JP
Associated scholarship(s):22/13551-4 - Simultaneous biopsy and lipid profile of embryo inner cell mass and trophectoderm, BE.EP.DR

Abstract

The aim of this work is to establish simultaneous biopsy and lipid profile of embryo inner cell mass and trofectoderm. For this purpose, oocytes recovery from slaughterhouse ovaries will be submitted to in vitro maturation and fertilization (D0). The presumptive zygotes will be cultured in vitro to produce the blastocysts. Experiment I: day 7 expanded blastocysts (XB, n = 45 / group) will be submitted to the Inner Cell Mass (ICM) and trofectoderm (TE) biopsy procedure. The biopsies will be appropriately stored for further evaluation (protein immunolocalization and lipid profile) and the biopsied embryos will return to the in vitro culture to developmental monitoring, separated as follows: Group 1: XB underwent biopsy of both ICM and TE; group 2: XB underwent only TE biopsy; group 3: non-biopsied XB. After 12 hours of culture, a morphological analysis and the re-expansion rate of the biopsied blastocysts (only embryos from groups 1 and 2) will be performed, followed by the investigation of apoptosis (TUNEL) and immunolocalization of proteins (CDX2, GATA6 and NANOG) in the blastocysts of all groups (groups 1 to 3) to evaluate the biopsy effect on the embryo development, apoptosis and differentiation of the different cell lines (trofectoderm, epiblast and hypoblast). Experiment II: biopsied ICM and TE cells (n = 15, groups 1 and 2) will be used for protein characterization of ICM and TE through the investigation of CDX2 protein. Experiment III: ICM and TE biopsies (groups 1 and 2) will be grouped in pools of 3 biopsies per replicate (n = 5 replicates/group) and submitted to lipid profile analysis by Multiple Reaction Monitoring (MRM-profiling) at an Omics Facility Lab. For statistical analysis, uni and multivariate models will be used in robust statistical software capable of evaluating a large amount of data and provide strong results to the hypotheses tested. The significance level of 5% (P <0.05) will be adopted. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
VALENTE, RONIELE SANTANA; MARSICO, THAMIRIS VIEIRA; SUDANO, MATEUS JOSE. Basic and applied features in the cryopreservation progress of bovine embryos. Animal Reproduction Science, v. 239, p. 8-pg., . (18/21496-8, 19/17796-9, 19/18415-9)
MARSICO, THAMIRIS VIEIRA; VALENTE, RONIELE SANTANA; ANNES, KELLY; OLIVEIRA, ANDRESSA MINOZZO; SILVA, MARA VIANA; SUDANO, MATEUS JOSE. Species-specific molecular differentiation of embryonic inner cell mass and trophectoderm: A systematic review. Animal Reproduction Science, v. 252, p. 11-pg., . (19/17796-9, 18/21496-8, 20/08826-9, 22/02139-5, 19/18415-9)
MARSICO, THAMIRIS VIEIRA; SILVA, MARA VIANA; VALENTE, RONIELE SANTANA; ANNES, KELLY; RISSI, VITOR BRAGA; GLANZNER, WERNER GIEHL; SUDANO, MATEUS JOSE. Unraveling the Consequences of Oxygen Imbalance on Early Embryo Development: Exploring Mitigation Strategies. ANIMALS, v. 13, n. 13, p. 18-pg., . (19/17796-9, 18/21496-8, 19/18415-9, 20/08826-9)

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