Coculture of porcine cumulus–oocyte complexes with porcine luteal cells during IVM: effect on oocyte maturation and embryo development
G. M. Teplitz A B , M. S. Lorenzo A B , A. Maruri B , P. R. Cruzans A B , M. C. Carou B and D. M. Lombardo B CA Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Godoy Cruz 2290 C1425TQB, Buenos Aires, Argentina.
B Universidad de Buenos Aires, Facultad de Ciencias Veterinarias, Instituto de Investigación y Tecnología en Reproducción Animal, Cátedra de Histología y Embriología, Chorroarín 280 C1427CWO, Buenos Aires, Argentina.
C Corresponding author. Email: dlombard@fvet.uba.ar
Reproduction, Fertility and Development 32(16) 1250-1259 https://doi.org/10.1071/RD20117
Submitted: 29 April 2020 Accepted: 12 September 2020 Published: 21 October 2020
Abstract
Coculture with somatic cells is an alternative to improve suboptimal in vitro culture conditions. In pigs, IVF is related to poor male pronuclear formation and high rates of polyspermy. The aim of this study was to assess the effect of a coculture system with porcine luteal cells (PLCs) on the IVM of porcine cumulus–oocyte complexes (COCs). Abattoir-derived ovaries were used to obtain PLCs and COCs. COCs were matured in vitro in TCM-199 with or without the addition of human menopausal gonadotrophin (hMG; C+hMG and C-hMG respectively), in coculture with PLCs from passage 1 (PLC-1) and in PLC-1 conditioned medium (CM). In the coculture system, nuclear maturation rates were significantly higher than in the C-hMG and CM groups, but similar to rates in the C+hMG group. In cumulus cells, PLC-1 coculture decreased viability, early apoptosis and necrosis, and increased late apoptosis compared with C+hMG. PLC-1 coculture also decreased reactive oxygen species levels in cumulus cells. After IVF, monospermic penetration and IVF efficiency increased in the PLC-1 group compared with the C+hMG group. After in vitro culture, higher blastocysts rates were observed in the PLC-1 group. This is the first report of a coculture system of COCs with PLCs. Our model could be an alternative for the conventional maturation medium plus gonadotrophins because of its lower rates of polyspermic penetration and higher blastocysts rates, key issues in porcine in vitro embryo production.
Keywords: coculture, embryo development, IVM, luteal cells, pig.
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