Synergistic Effect of Porcine Follicular Fluid and Dibutyryl Cyclic Adenosine Monophosphate on Development of Parthenogenetically Activated Oocytes from Pre-Pubertal Gilts

Abstract

This study investigated the effect of porcine follicular fluid (PFF) and dibutyryl cyclic adenosine monophosphate (dbcAMP) during in vitro maturation (IVM) of porcine oocytes on meiotic maturation, fertilization and embryo development, and compared the effect of supplementing the embryo culture media with PFF or foetal bovine serum (FBS) on embryo development. Oocytes from pre-pubertal gilts were IVM for 44 h, and parthenogenetically activated or in vitro-fertilized. Embryos were cultured in porcine zygote medium (PZM3) for 7 days. Cleavage and blastocyst rates were evaluated at 48 h and 7 days of culture. The supplementation of the IVM medium with 25% PFF and 1 mm dbcAMP for the first 22 h resulted in more (p < 0.05) embryos developing to the blastocyst stage as compared with the inclusion of dbcAMP alone. The dbcAMP + PFF combination increased (p < 0.05) the average number of nuclei per blastocyst as compared with either of these components alone or in its absence. A synergistic effect of dbcAMP + PFF during IVM was also reflected in the capacity of oocytes to regulate sperm penetration and prevent polyspermy, as twice as many oocytes from the control group were penetrated by more than one sperm as compared with those matured in the presence of both dbcAMP and PFF. The supplementation of PZM3 with 10% FBS from days 5 to 7 of culture significantly improved the total cell quantity in embryos derived either from control or dbcAMP + PFF matured oocytes. There was no effect on the total cell quantity when FBS was replaced by the same concentration of PFF. These studies showed that dbcAMP, PFF and FBS can improve both the quantity (57.3% vs 41.5%) and quality (74.8 vs 33.3 nuclei) of porcine blastocysts derived from oocytes recovered of pre-pubertal gilts.

Short-term Cultivation of Porcine Cumulus Cells Influences the Cyclin-dependent Kinase 4 (Cdk4) and Connexin 43 (Cx43) Protein Expression—A Real-time Cell Proliferation Approach

“…However, the molecular changes, visible as differential expression profiles of genes and proteins responsible for COC maturation, are still not fully recognized. It was shown in several studies that the achievement of a fully mature stage of oocytes is regulated via an appropriate communication between the oocyte and the surrounding cumulus-granulosa cells, and it is assumed that attainment of the MII stage by the oocyte is accompanied by CC expansion [8, 18, 19, 24, 25]. The mechanisms responsible for this process are not entirely known.…”

“…In order to provide optimal conditions for subsequent oocyte maturation and fertilization in vitro , the ovaries of each animal were placed in 5% fetal bovine serum solution (FBS; Sigma-Aldrich, St. Louis, MO, USA) in PBS [18, 19]. Thereafter, individual large follicles (>5 mm) were opened by puncturing with a 20-G needle connected to a 5-ml syringe in a sterile Petri dish, and COCs were recovered.…”

Phosphorylated histone H2A.x in porcine embryos produced by IVF and somatic cell nuclear transfer

“…Because pigs are physiologically similar to humans, there is growing interest in the use of these technologies to create special animal models for the study of physiopathological processes and xenotransplantation (Kues & Niemann 2004, Kuwaki et al 2005, Aigner et al 2010. However, the development of IVF and SCNT porcine embryos is influenced by different factors, including oocyte quality, polyspermy, abnormal cell cycle interactions, deficient epigenetic reprogramming, and altered gene expression (Abeydeera et al 2000, Bortvin et al 2003, Lucifero et al 2006, Niemann et al 2008, Nascimento et al 2010, Whitworth & Prather 2010, Yoshioka 2011.…”

Time- and Dose-Dependent Effects of 17 Beta-Estradiol on Short-Term, Real-Time Proliferation and Gene Expression in Porcine Granulosa Cells

“…To provide optimum conditions, the ovaries of each animal were placed in 5% fetal bovine serum (FBS; Sigma-Aldrich Co., St. Louis, MO, USA) in PBS [7, 8]. Granulosa cells were collected by aspirating the follicular fluid from single follicles using a 20 G needle.…”