Isolation and expansion of cytokeratin positive progenitor cells from adult murine pancreatic ducts expressing Pdx-1, Nestin, Sox9, MafA and hepatic nuclear factors.
BACKGROUND
Recent studies suggest that stem cells may represent a putative source for the generation of beta cells. However, the identity and characteristics of stem cells from adult pancreas and conditions for their large scale expansion are still poorly defined.


METHODS
DPC were isolated from adult pancreatic ducts of C57Bl/6 mice. Expression profile was investigated by PCR, FACS and immunohistochemistry.


RESULTS
DPC express a panel of stem cell associated markers such as Pdx-1, cytokeratin-19 (CK19), nestin, Sox9 together with the transcription factor MafA and hepatic nuclear factors HNF1β, HNF3β, HNF4α und HNF6. This gene expression profile is suggesting that DPC might be a promising tool for endocrine differentiation. After stimulation with picolinic acid and hypoxia, DPC expressed the endocrine differentiation marker Ngn3. Nevertheless, insulin production was not observed.


CONCLUSIONS
We here describe a protocol for the isolation end expansion of murine pancreatic ductal progenitor cells (DPC) displaying high self-renewal, spheroid- and colony-forming capacity. Further studies are required to elucidate the conditions for differentiation into mature pancreatic endocrine cell lineages.
IV Medical Department, Ludwig Maximilians University, Munich, Germany - ulrich.grabmaier@med.uni-muenchen.de.|I Medical Department, Ludwig Maximilians University, Munich, Germany - ulrich.grabmaier@med.uni-muenchen.de.
Ulrich
U
Grabmaier
IV Medical Department, Ludwig Maximilians University, Munich, Germany.
Isabelle
I
Rottstegge
IV Medical Department, Ludwig Maximilians University, Munich, Germany.
Monika
M
Offers
I Medical Department, Ludwig Maximilians University, Munich, Germany.|Department of Molecular Medicine, Max Planck Institute of Biochemistry, Munich, Germany.
Tilmann
T
Ziegler
I Medical Department, Ludwig Maximilians University, Munich, Germany.
Christoph
C
Brenner
I Medical Department, Ludwig Maximilians University, Munich, Germany.
Bruno C
BC
Huber
IV Medical Department, Ludwig Maximilians University, Munich, Germany.
Jochen
J
Seissler
D000818:Animals; D002460:Cell Line; D019070:Cell Lineage; D051537:Hepatocyte Nuclear Factor 1 / Q000235:genetics*; D018398:Homeodomain Proteins / Q000235:genetics*; D007633:Keratins / Q000378:metabolism*; D051473:Maf Transcription Factors, Large / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D064231:Nestin / Q000235:genetics*; D010183:Pancreatic Ducts / Q000166:cytology* / Q000378:metabolism*; D055758:SOX9 Transcription Factor / Q000235:genetics*; D013234:Stem Cells / Q000378:metabolism*; D015534:Trans-Activators / Q000235:genetics*
D018398:Homeodomain Proteins; D051473:Maf Transcription Factors, Large; C485160:Mafa protein, mouse; C577485:Nes protein, mouse; D064231:Nestin; D055758:SOX9 Transcription Factor; C527933:Sox9 protein, mouse; D015534:Trans-Activators; C084010:pancreatic and duodenal homeobox 1 protein; D051537:Hepatocyte Nuclear Factor 1; D007633:Keratins