Neogenisis of nestin positive islet precursor cells from rat

 

M. Subramaniam, E. Brown, P.G. Shiels, D. Deardon

University of Glasgow

Division of Cancer Sciences and Molecular Pathology

Department of Surgery, Western Infirmary Glasgow

Scotland, UK

 

Intoduction:

World wide five million people suffer from type 1 diabetes and despite insulin therapy, late complications such as retinopathy, nephropathy, neuropathy etc are not uncommon.

Islet transplantation or allografting of islet precursor cells, has been suggested as a possible means of circumventing these problems. Current islet transplantation procedures are disadvantaged by a requirement of high numbers of islets (~million islet cells) and a high percentage loss of functional cells post transplant. Islets derived from progenitor cells is one solution to this dilemma. We have investigated the possibility of deriving islets from progenitor cells isolated from pancreatic ducts and have successfully achieved neogenesis of nestin positive islet progenitor cells from rat pancreatic ductal cells. 

 

Materials and Methods:

Pancreatic ducts were dissected from the pancreas of a six month old male AS rat. The ducts were micro dissected and incubated in type 5 collagenase for 20 minutes.  The minced pancreatic ducts were incubated at 370 C and 5% CO2 in culture medium.

 

Results:

Incubation of pancreatic ductal tissue resulted in the isolation of slow growing elongated cells (Fig1). These were assessed for expression of nestin, as a primary progenitor cell marker, using RT-PCR (fig2). They were PDX-1 negative and the phenotypic appearance of these cells remained unchanged despite passage. We were able to differentiate these cells into islet producing clusters and neuronal cells.

 

Conclusion: We hope to produce islet cells in the near future.