Selectively Inhibition of MAPK pathway showed positive effects on Porcine embryo-derived stem cells pluripotency

Expanded pluripotent stem cells (EPSCs), derived from pre-implantation embryos in specific culture conditions, can contribute to both embryonic and extraembryonic lineages. Human and mouse EPSCs have been successfully established in the LCDM condition, however, porcine embryo-derived stem cells derived in the LCDM system lack expanded pluripotency. To investigate the species-specific molecular mechanism of porcine embryo-derived stem cells (pLCDM) in LCDM condition, we compared the biological characteristics of pLCDM and porcine expanded potency stem cells (pEPSCs). The results revealed that pLCDM exhibited unique pluripotent traits and limited embryonic lineage differentiation potential, with a strong inclination towards extra-embryonic lineage differentiation. Subsequent investigation showed that (S)-(+) -dimethindene maleate (DIM) could completely inhibit the MAPK signaling pathway in pLCDM, and there were significant differences in the activity of the P38, JNK and ERK branches of the MAPK signaling pathway between pLCDM and pEPSC. Screening key molecular target inhibitors in three branches identified the B-RAF inhibitor SB590885 (SB59) as enhancing porcine embryonic stem cell pluripotency and facilitating lineage differentiation. Further analysis revealed a significant correlation between the expression level of phosphorylated ERK (p-ERK) and the developmental potential of porcine embryo-derived stem cells. These findings contribute valuable insights into the understanding of regulatory mechanism of porcine pluripotency.