Acta Pharmacologica Sinica (2009) 30: 1289–1296; doi: 10.1038/aps.2009.115; published online 24 August 2009

Aim: The feasibility of inducing endocrine pancreatic differentiation of embryonic stem (ES) cells has been well documented.  However, whether ES cells possess the potential for exocrine pancreatic differentiation requires further exploration.  Here, we investigated whether sodium butyrate and glucocorticoids were conducive to the exocrine pancreatic differentiation of ES cells.

Methods: E14 mouse ES cells were cultured in suspension to form embryoid bodies (EBs).  These EBs were cultured in differentiating medium containing varying concentrations of sodium butyrate.  The effects of activinA and dexamethasone (Dex) on exocrine differentiation were also explored.  Finally, the combination of sodium butyrate, activinA, and Dex was used to promote the differentiation of exocrine pancreatic cells.  Specific exocrine pancreatic gene expression was detected by reverse transcription polymerase chain reaction (RT-PCR) and amylase expression was examined by immunofluorescence staining.  Flow cytometry analysis was also performed to determine the percentage of amylase-positive cells after the treatment with activinA, sodium butyrate, and Dex.

Results: Exposure of ES cells to 1 mmol/L sodium butyrate for 5 days promoted exocrine pancreatic gene expression.  Further combination with Dex and other pancreatic-inducing factors, such as activinA, significantly enhanced the mRNA and protein levels of exocrine pancreatic markers.  Additionally, flow cytometry revealed that approximately 17% of the final differentiated cells were amylase-positive. 

Conclusion: These data indicate that the exocrine pancreatic differentiation of ES cells can be induced by activinA, sodium butyrate, and Dex, providing a potential tool for studying pancreatic differentiation and pancreas-related diseases.

Keywords: embryonic stem cells; sodium butyrate; dexamethasone; differentiation; exocrine pancreatic cells