The Effects of Basal Medium and Small Molecules on the Efficiency of Porcine Embryonic Stem Cell-like Cell Lines Establishment

Abstract: Swine is considered as one of the most important candidates for human disease models， and establishment of bona fide porcine embryonic stem cells (pESC) will facilitate making disesase models. Culture medium and signalling pathway are two main factors that affect the establishment of porcine embryonic stem cells. In this study， we investigated the effects of basal medium (KO-DMEM and DMEM/F12) and small molecular inhibitors (PD0325901 and CHIR99021， 2i) on the efficiency of pESC establishment. The results showed that the rates of attached embryos and primary colonies in KO-DMEM basal medium were significantly higher than those in DMEM/F12 basal medium when IVF embryos of 6 days were seeded， and the rates were significantly decreased when embryos seeded in both of the two medium with addition of 2i. pESC-like cell lines were derived from KODMEM basal medium. These cells were alkaline phosphatase (AP) positive and had normal karyotypes， expressed pluripotent markers of Oct4， Sox2 and Nanog， but did not express Cdx2. The cell lines could be genetically modified without losing of the pluripotency. These results indicated that KO-DMEM basal medium could support pESClike cells derivation and 2i had negative effects on embryos attachment and primary colonies formation. This study provided important evidence for selection of basal medium and predicting of pluripotency related cell signalling in pESC establishment.

CSTR: 32200.14.cjcb.2015.05.0004