Abstract: The pig, as a biological analog of human in multiple biomedical research due to its more conserved physiological, metabolic, and genetic characteristics to human than lower mammals such as rodents, has long been considered as a useful model for studying mammalian embryonic development and human disease. However, it has been challenging to establish pluripotent stem cells from pig preimplantation embryos. Here we report the derivation of Naïve-Like Porcine Pluripotent Stem Cell (N-L pPSC) line from porcine Expanded Potential Stem Cell (pEPSC) and pre-implantation embryo through the optimization of culture condition with PD0325901 included, the key inhibitor for capturing and maintenance of human and mouse naïve pluripotent stem cells. Our newly established N-L pPSCs are robust in culture with high expression of typical pluripotency factors such as OCT4, SOX2 and NANOG. We have also observed remarkable upregulation of the naïve pluripotency markers including TFAP2C and KLF4 compared to pEPSCs indicating the naïve state transition. Moreover, N-L pPSCs appear to be highly efficient in deriving porcine trophoblast stem cells (pTSCs) and porcine blastocyst-like structures (blastoids) indicating retaining an extraembryonic lineage differentiation potential. In vitro differentiation assays confirmed that N-L pPSCs formed three-dimensional embryoid bodies, spontaneously differentiated into cell types from all three germ layers as well as trophoblasts. The high differentiation potency and molecular features indicate our derivation of porcine naïve-like pluripotent stem cells.
Funding Source: This project is supported by Health@InnoHK, Innovation Technology Commission, HKSAR
