Follicle mural granulosa cells encapsulated in sodium alginate gel improve developmental competence of porcine oocytes of in vitro maturation

Abstract

The maturation of oocytes has an important impact on the subsequent development of the embryo. However, during the in vitro maturation (IVM) of oocytes, oocytes are separated from the follicular environment, resulting in a low maturation rate of oocytes in vitro. In order to improve maturation rate of IVM of porcine oocytes, this study was conducted to investigate using sodium alginate (SA) to encapsulate porcine mural granulosa cells (MGs) to develop an SA three-dimensional (3D) co-culture system for IVM of porcine oocytes. And, gene expression, reactive oxygen species (ROS), ATP level, mitochondrial membrane potential, parthenogenetic activation development results of cultured oocytes, and as well as ROS and glutathione (GSH) levels in cumulus granulosa cells (CGs) were detected. Our results showed that the maturation rate of the SA 3D co-culture group was 85.41 %, that of the negative control (NC) group was 79.24 %, and that of the MGs co-culture group was 81.62 %. In SA 3D co-culture group, mitochondrial membrane potential level of oocytes was 1.6, ROS level was 19 and the ATP level was 1.7. While in NC group, mitochondrial membrane potential level of oocytes was 1.2, the ROS level was 52, and the ATP level was 0.4. The ROS level in the CGs of SA 3D co-culture group decreased by 1.5 times, and the glutathione content increased by 2.3 times. In the SA 3D co-culture group, GDF9 gene expression level was 2.0, and BMP15 gene expression level was 1.2. While in NC group, GDF9 gene expression level was 0.7, and BMP15 gene expression level was 0.6. The blastocyst rate in the SA 3D co-culture group was 41.4 %, and that in the NC group was 36.6 %. In conclusion, encapsulating MGs in SA gel and co-culturing them with porcine oocytes in 3D during IVM can improve the developmental potential of oocytes. This result will provide an important reference for improving the methods of in vitro maturation of oocytes.