Evaluation of mass spectrometry for bovine in vitro production media and fetal calf serum quality control

• Grant number: 07/01400-1
• Support Opportunities: Research Grants - Innovative Research in Small Business - PIPE
• Start date: October 01, 2007
• End date: April 30, 2008
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Christina Ramires Ferreira
• Grantee: Christina Ramires Ferreira
• Company: In Vitro Brasil Ltda
• City: Mogi Mirim
• Associated researchers: Gustavo Henrique Martins Ferreira de Souza ; Marcos Nogueira Eberlin

Abstract

In Latin America, especially Brazil, is responsible for almost 50% of all in vitro-derived bovine embryos produced in the world. Quality control (QC) procedures for bovine embryo in vitro production (IVP) media include pH and osmolarity checking. In some situations, embryo development rate from slaughterhouse-derived oocytes is evaluated. IVP media are complex mixtures of salts, proteins, hormones, carbohydrates and vitamins, which may have their composition altered due continuous reagent handling, to the storage conditions and even due to technical failure preparation. Consequences may involve embryo development rate decrease or failure, leading to economical 1055 in commercial IVP systems. Mass spectrometry (MS) is an analytical and highly sensible tool, used for unknown compound identification; organic, inorganic and complex molecule analysis; also to find target molecules structures and chemical properties. MS technique allows target compounds to be identified inside complex mixtures in extremely low concentrations (femtomols). Matrix-Assisted Laser Desorption/Ionization (MALDI) technique was developed to promote large polypeptide and protein ionization, but it was further employed for glycoproteins, oligonucleotides and complex carbohydrates analysis. One of MALDI- Time of Flight (TOF) great advantages concern in fact the ionization process causes un-feasible or a very low molecular fragmentation, allowing molecular ion identification even in a complex mixture. This project proposes the use of mass spectrometry with MALDI-TOF ionization source as a technique for bovine IVP media QC. Mass/charge ratio (m/z) profiles will be obtained from (1) in vitro maturation, (2) in vitro fertilization, (3) in vitro culture, and (4) embryo transportation media. Detected ions will be compared considering the storage time (0, 3, 7, 10 and 14 days) at 4oC. Seven batches of fetal calf serum (FCS), the most critical supplement in bovine IVP, will also be evaluated using MALDI-TOF. Results regarding MALDI-TOF analyzed media will be related to embryo development data obtained from slaughterhouse-derived oocytes. MALDI-TOF will be used to detect 16 composition changes in the IVP media. After providing spectra from ion intensity differences related to the media storage period, it will be possible media quality markers identification and to establish a wide and fast bovine IVP media QC and check interval. Regarding to the FCS, an ions profile related to the best development rates will be defined. Initial experiments, reported in this project, indicated that MALDI-TOF allowed consistent spectra acquisition, with medium to high intensity of some ions, without previous sample preparation. Therefore, this project aims to develop a aim pie, fast and direct method, efficient for providing commercial bovine IVP media QC. In Vitro Brasil Ltda company produces large amount of bovine IVP media every week and sells them for 2 branches and to partner laboratories. The possibility to measure media quality would provide riskless media distribution system, since this product influences IVP success in great extent. (AU)

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)

FERREIRA, CHRISTINA RAMIRES; MARTINS FERREIRA SOUZA, GUSTAVO HENRIQUE; RICCIO, MARIA FRANCESCA; CATHARINO, RODRIGO RAMOS; FORTES PONTES, JOSE HENRIQUE; BASSO, ANDREA CRISTINA; ERENO JUNIOR, JOSE CARLOS; PERECIN, FELIPE; EBERLIN, MARCOS NOGUEIRA. Mass spectrometry fingerprinting of media used for in vitro production of bovine embryos. RAPID COMMUNICATIONS IN MASS SPECTROMETRY, v. 23, n. 9, p. 1313-1320, MAY 2009. (07/01400-1)