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Pancreatic progenitor cells and methods for isolating the same

a pancreatic progenitor cell and pancreatic stem cell technology, applied in the field of models for identifying and isolating pancreatic stem cells, can solve the problems of pathological consequences of targeted cell loss or aberrant cell growth and developmen

Inactive Publication Date: 2007-06-28
SARVETNICK NORA +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

"The invention provides animal models for identifying pancreatic stem / progenitor cells and a common stem / progenitor cell that gives rise to liver cells and pancreatic cells. The animals show no pathology, hyperglycemia, or hypoglycemia associated with the KGF expression. The invention also provides methods of making and proliferating the transgenic cells and methods of using the transgenic cells. The invention provides a pancreactic duct culture useful for identifying a common stem / progenitor cell in pancreatic islets. The invention also provides a method for using the homeobox protein PDX-1 as a marker for identifying a common stem / progenitor cell."

Problems solved by technology

A healthy adult pancreas is usually a developmentally stable organ, but a targeted cell loss or aberrant cell growth and development have pathological consequences.

Method used

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Examples

Experimental program
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Effect test

example 1

Expression of KGF in the Islets of Langerhans Leads to Pancreatic Hepatocyte Generation and Duct Cell Proliferation

[0079] To better understand the role of KGF in pancreatic development, a model system was established in which the influence of localized KGF expression on the growth and development of the pancreas can be assessed. A transgenic mouse model was constructed in which expression of the murine KGF-coding polynucleotide controlled by the human insulin promoter, resulting in KGF expression within beta cells in the islets of Langerhans. Ectopic KGF expression resulted in several changes to the pancreas, as described below. The ins-KGF mouse therefore provides a valuable system for studies designed to enhance understanding of pancreatic growth and development.

[0080] Generation of ins-KGF transgenic mice To generate ins-KGF transgenic mice, the 585 base pair (bp) KGF cDNA was obtained by reverse transcriptase polymerase chain reaction (RT-PCR) of mRNA from mouse salivary gland...

example 2

Transgenic Expression of EGF in Pancreatic Beta Cells Results in Substantial Morphological Changes

[0091] EGF was ectopically expressed in transgenic mice using a human insulin promoter. The beta cell-targeted expression of EGF resulted in significant morphological changes, including cellular proliferation and disorganized islet of Langerhans growth. The mice were normnoglycemic. Interestingly, insulin-producing beta cells were found in some of the ducts of older ins-EGF transgenic mice. This EXAMPLE therefore shows that both EGF can affect pancreatic development and growth.

[0092] Generation of ins-EGF transgenic mice To determine the influence of EGF expression in the beta cells of the pancreas, transgenic mice expressing murine EGF under control of the human insulin promoter were generated (ins-EGF). The 280-bp EGF cDNA was used to generate the ins-EGF transgenic mouse. The cDNA was cloned into a vector containing the human insulin promoter and the hepatitis b 3′ untranslated pol...

example 3

Transgenic Expression of EGF and KGF in Pancreatic Beta Cells Results in Substantial Morphological Changes

[0101] Two important growth factors for pancreatic development, EGF and KGF, were ectopically expressed in transgenic mice using the human insulin promoter. The beta cell-targeted expression of either EGF or KGF resulted in significant morphological changes, including cellular proliferation and disorganized islet of Langerhans growth In both cases, the mice were normoglycemic. Intercrossing the individual ins-EGF and ins-KGF transgenic mice to generate Ins EGF x KGF mice resulted in transgenic mice with more profound changes in pancreatic morphology than was seen for either growth factor alone. Proliferation of pancreatic cells was also observed in the double transgenic mouse, as was extensive intra-islet fibrosis, which was found to increase in severity with time. Interestingly, insulin-producing beta cells were found in some of the ducts of older ins-EGF and ins-EGF x KGF tra...

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Abstract

The invention provides animal models, where the ectopic expression of KGF, EGF, or both is under the control of a pancreas-specific promoter, e.g., the insulin promoter. The expression of KGF in the ins-KGF pancreatic islets of Langerhans results in enlarged islets, with substantial proliferation of duct cells within the islet mass, and the presence of albumin and alpha-fetoprotein-producing hepatocytes in the islets of the ins-KGF pancreata. The compositions and methods disclosed are useful for identifying and isolating pancreatic stem / progenitor cells, including a common stem / progenitor to liver cells and pancreatic cells.

Description

TECHNICAL FIELD [0001] This invention relates to generally to models for identifying and isolating pancreatic stem cells, and in one embodiment, to models for identifying and isolating a common stem / progenitor cell that gives rise to hepatic cells and pancreatic cells. BACKGROUND OF THE INVENTION [0002] A healthy adult pancreas is usually a developmentally stable organ, but a targeted cell loss or aberrant cell growth and development have pathological consequences. For example, in human Insulin Dependent Diabetes Mellitus (IDDM), autoimmune mechanisms cause the selective and permanent destruction of the insulin-producing beta cells in the islets of Langerhans. The lost cells are not restored in vivo, although Sarvetnick et al. Cell 52:773-782 (1988) have shown that beta cells have the potential to regenerate. On the other hand, the proliferation of duct cells can contribute to pancreatic disease pathologies, such as chronic pancreatitis, pancreatic cancer, and cystic fibrosis. Pancr...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): G01N33/567A01K67/027A61K38/00A61K38/18A61K39/00A61K39/395A61K48/00A61P1/16A61P1/18C07K16/18C12N1/00C12N5/071C12N5/074C12N5/10C12N15/09C12N15/85C12Q1/02C12Q1/68G01N33/53
CPCA01K67/0271A01K67/0275A01K2217/05A01K2227/105A01K2267/03A01K2267/0325A61K38/1808A61K38/1825A61K39/00A61K48/00C07K16/18C07K2319/30C12N5/0672C12N5/0676C12N5/0678C12N15/8509C12N2501/11C12N2501/117C12N2503/00C12N2517/02C12N2830/008G01N33/5005G01N33/566A61P1/16A61P1/18A61P3/00
Inventor SARVETNICK, NORAKRAKOWSKI, MICHELLE L.KRITZIK, MARCIE R.
Owner SARVETNICK NORA
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