Abstract
Background
The specific precursor cells derived from embryonic stem cells (ESCs) after induced differentiation offers great potential for repairing damaged pancreas. Therefore, it is of crucial importance to know whether pancreatic precursor cells with Pdx1+ could be induced to differentiate into pancreatic-like cells in vitro as well as the probable mechanism underlying.
Methods
In this study, mouse ESCs (ES-E14TG2a) were divided into the blank control group (ESC), the blank vector group (Pdx1− ESC), and the Pdx1 lentiviral vector group (Pdx1+ ESC). We constructed lentiviral vectors with overexpressed Pdx1 in Tet-on system and screened the stably transfected cell lines after transfection. CXCR4+ (C-X-C chemokine receptor type 4) DE cells of the three groups were seeded (day zero [d0]) after being sorted with immune beads, and expression of Pdx1 was induced on the second day (d1). Then, markers for the differentiation of Pdx1+ pancreatic precursor cells and molecules in the Notch pathway were detected at d3, d7, d10, and, d14.
Results
We found that expressions of Ptf1a, CK19, and amylase increased at d3 and d7, Neuro D1 increased at d10 and d14, Pax6 and insulin increased at d14, as well as Notch1, Notch2, Hes1, and Hes5 increased at d3 and thereafter declined at d14 in the Pdx1+ ESC group. These expressions were significantly higher than those in the ESC group and Pdx1− ESC group, but no marked differences were observed between the ESC group and Pdx1− ESC group.
Conclusions
Our study indicates that Pdx1+ pancreatic precursor cells can differentiate into pancreatic-like cells in which the Notch pathway plays important roles.
908. Lentiviral Vector-Mediated Transduction in Murine SMA Embryonic Stem Cells Derived from Spinal Muscular Atrophy (SMAI) Mice