Comparison of different cell culture medium for in vitro production of bovine embryos in the absence of fetal calf serum

• Grant number: 11/13463-3
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: October 01, 2011
• End date: September 30, 2012
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Fernanda da Cruz Landim
• Grantee: Henrique Pedro Dias
• Host Institution: Faculdade de Medicina Veterinária e Zootecnia (FMVZ). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

The culture media utilized for in vitro embryo production affects development after fecundation, interfering with embryo metabolism, gene expression, and pregnancy rates. Bovine embryos are, normally cultured in media supplemented with fetal calf serum (FCS). The serum contains several beneficial substances as growth factors, energetic substrates, aminoacids, vitamins, and antioxidants. The presence of the FCS promotes the computation of the morula and the expansion of the blastocyst. However, this compound has a biphasic effect on embryo development since its presence just after fertilization inhibits the first cleavage. Moreover, the serum is considered an indefinite product, interfering with the repeatability of the results obtained. Also, the utilization of FCS has been associated with lipid accumulation in vitro-produced embryos. The use of chemically defined media, without the addition of FCS, is an alternative for the in vitro production of embryos without the presence of animal additives aiming at the improvement of cryosurvival. So the goal of the present experiment is to test 3 different culture media to produce bovine embryos in vitro in culture systems in absence of FCS. The present study proposes the use of media normally used for embryonic stem cell cultures, such as DMEM and Alfa-MEM, in association with F12. Those media are not routinely used for embryo culture and their use will be compared with Sofia. As a replacement for FCS 10mg/mL of BSA Will is added to the media. This concentration was tested in a previous experiment performed by the same author with good embryo production.(AU)