Stimulation of cAMP signalling allows isolation of clonal pancreatic precursor cells from adult mouse pancreas.
OBJECTIVE
Duct cells of the pancreas are thought to include latent progenitors of islet endocrine cells that can be induced to differentiate by appropriate morphogens. Here we developed a method for isolating pancreatic ductal epithelial cells from adult mice that overcomes the shortcomings of previous methods.


METHODS
Pancreatic ductal cells were grown in serum-free DMEM/F12 medium in the presence of cholera toxin or 8-bromo-cyclic adenosine monophosphate, which is known to be an intracellular cAMP generator. Single cell cloning was performed by limiting dilution in serum-free medium.


RESULTS
The isolated clonal cells expressed high levels of cytokeratin and Ipf1 (formerly known as Pdx-1). Adenovirus-mediated expression of ngn3 (also known as Neurog3) and Ptf1a in these cells induced expression of insulin and somatostatin, and of carboxypeptidase A, respectively. Furthermore, albumin production was induced by dexamethasone or by long-term culture in serum-containing medium.


CONCLUSIONS
Stimulation of the cAMP-dependent signalling allowed us to isolate clonal pancreatic ductal cells from adult mice. These cells are able to partially differentiate into endocrine cells, exocrine cells and hepatocyte-like cells and are therefore considered to have the characteristics of endodermal progenitor cells.
Yamamoto
T
T
Division of Stem Cell Regulation Research, G6, Osaka University Graduate School of Medicine, 2-2 Yamadaoka, Suita, Osaka, 565-0871, Japan.
Yamato
E
E
Taniguchi
H
H
Shimoda
M
M
Tashiro
F
F
Hosoi
M
M
Sato
T
T
Fujii
S
S
Miyazaki
J-I
JI
D000818:Animals; D018929:Cell Culture Techniques; D002460:Cell Line; D016895:Culture Media, Serum-Free; D000242:Cyclic AMP / Q000502:physiology*; D017931:DNA Primers; D047108:Embryonic Development; D004847:Epithelial Cells / Q000166:cytology / Q000502:physiology; D005786:Gene Expression Regulation; D005952:Glucose-6-Phosphatase / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D010183:Pancreatic Ducts / Q000166:cytology / Q000196:embryology / Q000502:physiology* / Q000648:ultrastructure; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000502:physiology*
D016895:Culture Media, Serum-Free; D017931:DNA Primers; D012333:RNA, Messenger; D000242:Cyclic AMP; D005952:Glucose-6-Phosphatase