earticle

영어

Recent decade, the studies for generation of chimeric organs have been advanced remarkably with the production of chimeric animals using by pluripotent stem cells (PSCs). However, the distribution rate of injected cells into an embryo is still low and it is considered as one of the major reasons of low efficiency of chimera production. The purpose of this study is to investigate the effect of growth differentiation factor 8 (GDF8) on porcine chimeric embryo development. In first, we produced mCherry-marked porcine iPSCs (mCh-iPSCs). The mCh-iPSCs were injected into 8 cells or morula stage of in vitro fertilized (IVF) embryos at day 2 under a microscope with the micromanipulator. These chimeric embryos were then cultured porcine embryonic stem cell medium (DEME F10/low glucose with 15% FBS) for 18hr and then transferred to fresh porcine zygote medium 5 (PZM5 with or without 150 pg/mL of GDF8). We investigated the effect of supplemented GDF8 during in vitro culture (IVC) of these chimeric embryos on their cleavage patterns, blastocyst formation ratio, and injected cells’ distributions. Data were analyzed by ANOVA followed by Tukey’s range test using SPSS (Statistical Package for Social Science). After day 5 of in vitro chimeric embryo culture, the GDF8 supplement group was shown significantly higher Using the chimeric embryos of the blastocyst stage, we evaluated distributions of the injected cells by immune staining of SOX2 as porcine inner cell mass (ICM) marker. The GDF8 supplement group was shown significantly increased mCherry and SOX2 per SOX2 expressing cells ratio than control (49.2% VS 20.7, respectively). In conclusion, the supplementation of GDF8 during IVC of porcine chimeric embryos improved embryonic developmental competence and enhanced the distributions of injected cells into ICM of chimeric embryos at the pre-implantation stage.