CULTURE AND MULTIPLICATION OF PRIMORDIAL GERM CELLS IN CHICKEN EMBRYO CHORIOALLANTOIC MEMBRANES

• Grant number: 24/12988-5
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: October 01, 2024
• End date: September 30, 2025
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Ricardo Jose Garcia Pereira
• Grantee: Matheus Moraes Azevedo
• Host Institution: Faculdade de Medicina Veterinária e Zootecnia (FMVZ). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Primordial germ cells (PGCs) are the only cells capable of passing genetic information through many generations. This characteristic is attributed to their potential for self-renewal and differentiation, enabling the production of millions of sperm or oocytes. Harnessing PGCs offer promising methods for preserving genetic resources through intra- and interspecific transplants. However, these methods are currently limited by challenges in maintaining and multiplying PGCs in vitro, as well as the associated laboratory costs. Therefore, this project aims to evaluate the efficiency of culturing chicken PGCs on chorioallantoic membranes as an alternative low-cost method. Chicken PGCs will be collected from the bloodstream of 2 to 3-day-old embryos and cultured in vitro to establish a lineage with exponential growth. Subsequently, lineages reaching a minimum density of 150,000 cells/ ml will be diluted and injected (approximately 10,000 cells /10 ¼L) onto the chorioallantoic membrane of chicken embryos. Simultaneously, the same number of cells (10,000 cells/ well) will be cultured in 48-well plates as controls. Growth rate, viability, and expression of germ cell markers (SSEA-1, VASA, and EMA-1) will be evaluated after 10 days of incubation. We hypothesize that this experiment will develop a simple and cost-effective protocol for culturing PGCs in eggs, maintaining their viability and undifferentiated state.