Abstract
BackgroundIt has been suggested that the ectopic expression of PDX1, a dominant pancreatic transcription factor, plays a critical role in the developmental programming of the pancreas even from cells of unrelated tissues such as keratinocytes and amniotic fluid stem cells. In this study we have chosen to drive pancreatic development in human amnion epithelial cells by inducing endogenous PDX1 expression. Further, we have investigated the role of Epidermal Growth Factor (EGF) and Poly-L-Ornithine (PLO) on this differentiation process.ResultsHuman amnion epithelial cells expressed high levels of endogenous PDX1 upon transduction with an adenoviral vector expressing murine Pdx1. Other markers of various stages of pancreatic differentiation such as NKX6.1, SOX17, RFX6, FOXA2, CFTR, NEUROD1, PAX4 and PPY were also expressed upon Pdx1 transduction.Although initial expression of pancreatic progenitor markers was higher in culture conditions lacking EGF, for a sustained and increased expression EGF was required. Culture on PLO further increased the positive impact of EGF.ConclusionPancreatic marker expression subsequent to mPdx1 transduction suggests that this approach may facilitate the in vitro differentiation of hAECs into cells of the endocrine pancreas. This result may have important implications in diabetes therapy.Electronic supplementary materialThe online version of this article (doi:10.1186/s12861-016-0108-y) contains supplementary material, which is available to authorized users.
Highlights
It has been suggested that the ectopic expression of PDX1, a dominant pancreatic transcription factor, plays a critical role in the developmental programming of the pancreas even from cells of unrelated tissues such as keratinocytes and amniotic fluid stem cells
We attempted to initiate the process of pancreatic differentiation of human amnion epithelial cells (hAECs) by transient transduction of mouse Pdx1 (mPdx1), a critical factor in the development of the pancreas
Effect of Epidermal Growth Factor (EGF) on mPdx1 transduction Since previous reports have suggested that an environment lacking EGF drives the pancreatic differentiation process forward, we investigated if this is the case for hAECs transduced with mPdx1
Summary
It has been suggested that the ectopic expression of PDX1, a dominant pancreatic transcription factor, plays a critical role in the developmental programming of the pancreas even from cells of unrelated tissues such as keratinocytes and amniotic fluid stem cells. In this study we have chosen to drive pancreatic development in human amnion epithelial cells by inducing endogenous PDX1 expression. With current treatment strategies being inadequate in addressing the complications associated with the disease [2], an alternative cellbased therapy approach is urgently needed. Attempts have been made to differentiate amnion epithelial cells (AECs), placenta-derived multipotent progenitor cells and amniotic fluid stem cells down the pancreatic lineage [5,6,7,8,9]. The present study aims to bridge this gap in current research
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