Culture and propagation of spermatogonial stem cells using chorioallantoic membrane of chicken embryos

• Grant number: 17/24534-5
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: May 01, 2018
• End date: April 30, 2019
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Ricardo Jose Garcia Pereira
• Grantee: Brian Matayoshi Kanashiro
• Host Institution: Faculdade de Medicina Veterinária e Zootecnia (FMVZ). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Spermatogonial stem cells (SSCs) are the only male cells capable of transmitting genetic information over several generations. This characteristic is linked to their self-renewal and differentiation potential, and their ability to generate millions of spermatozoa. Thus, SSCs have produced promissing techniques for animal germplasm conservation through intra and interspecific transplantation. Nevertheless, the use of SSCs is limited by problems regarding their in vitro maintainance and propagation, and excessive lab costs. Therefore, we aim to assess the efficiency of SSCs cultivation in chorioallantoic membranes (CAM) of chicken embryos, with the purpose of creating an alternative and low cost method for the SSC culture. For such, quail testis will be dissociate and a suspension of SSC will be diluted and placed (about 1 x 106 cells/ 50µl) on the CAM of chicken embryos. Growth rate, viability, and cell differentiation will be monitored every 48 hours making a total of 192 hours (i.e. 8 days after inoculation). We expect in this study to develop a practical protocol for SSC culture in recipient eggs, capable of keeping viable cells in a undifferentiated state at low costs.