EFFECT OF THE COCULTURE OF PORCINE LUTEAL CELLS AND PORCINE CUMULUS-OOCYTE COMPLEXES ON CORTICAL REACTION, IN VITRO FERTILIZATION AND EMBRYO DEVELOPMENT

TEPLITZ, G.; LORENZO, MS.; CRUZANS, PR.; LUCHETTI, CG.; GHERSA, J; LOMBARDO, DM.

Ciudad Autónoma de Buenos Aires

Otro; IV REUNIÓN CONJUNTA DE SOCIEDADES DE BIOLOGÍA DE LA REPÚBLICA ARGENTINA; 2020

Sociedad Argentina de Biología

In pigs, in vitro fertilization (IVF) is related to high rates of polyspermy and in vitro embryo production (IVP) is still an inefficient biotechnology. Coculture with somatic cells is an alternative to improve suboptimal in vitro culture conditions. The aim of this study was to evaluate the effect of coculture of porcine luteal cells (PLC) and cumulus-oocyte complexes (COC) during in vitro maturation (MIV) on the cortical reaction (CR), IVF and embryo development. Slaughterhouse ovaries were used to obtain PLC and COC. Selected COC were matured in vitro for 44 h in TCM-199 supplemented with human menopausal gonadotropin (control) or in coculture with PLC. The IVF was performed with refrigerated sperm in 100 μL drops of TCM-199 supplemented medium for 4 h. Following IVF, suspected zygotes were stained with Hoechst 33342, and sperm penetration, monospermic penetration (monospermic / penetrated), and IVF efficiency (monospermic / total oocytes) were assessed. Cortical reaction (cortical granule distribution) was evaluated with FITC-PNA staining after IVF. For embryo development, suspected zygotes were washed and cultured in PZM at 39 ° C, 7% O2, 5% CO2. Cleavage rates were determined on day 2 and blastocysts rates on day 7. More cortical granules were observed in the area immediately below the oolemma, and the area outside the zona pellucida was more stained with PNA in the coculture group (p