Endocrine pancreas differentiation of adipose tissue-derived stromal cells and uses thereof

An adipose tissue and stromal cell technology, applied in the direction of bone/connective tissue cells, cells modified by introducing foreign genetic material, genetically modified cells, etc., can solve problems such as hepatic glucose decline

Inactive Publication Date: 2005-03-16
ARTECEL SCIENCE INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Circulating levels of PP also act on the liver and lead to a decrease in glycogen production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0161] In vitro induction method

[0162] Adipose-derived hepatocytes were isolated from liposuction waste material as described (Sen et al., 2001, J. Cell Biochem. 81:312-319). These cells were continued in culture in the presence (but not limited to) of the following media: Neurobasal supplemented with or without fetal bovine serum (FBS) TM (InVitrogen), N2, B27 (InVitrogen), or basic fibroblast growth factor (bFGF). Glucose levels in the medium were adjusted. Cells were grown at various densities and fed at 3-6 day intervals. Most preferably, they are present at about 1000 to about 500,000 cells / cm 2 planting density.

[0163]During culture, conditioned medium was assayed for the endocrine pancreatic hormones insulin (American Laboratory Products), glucagon, somatostatin, and pancreatic polypeptide (Peninsula Labs Inc) using commercially available radioimmunoassays or enzyme-linked immunosorbent assays. analyze.

[0164] Expression of phenotypic markers associated wit...

Embodiment 2

[0166] gene therapy method

[0167] The method includes inserting and expressing any gene that results in the induction of differentiation of the adult stem cells into cells expressing at least one genotypic or phenotypic characteristic of pancreatic cells. These genes may include, but are not limited to, controlled expression of the transcription factors HNF3β, Isl-1, Brain-4, Pax-6, Pax-4, Beta2 / NeuroD, PDX-1, Nkx6.2, Nkx2.2, and Ngn-3 . Potential methods of introducing nucleic acid into cells include, but are not limited to, electroporation, calcium phosphate, retrovirus, adenovirus, or lipid-mediated delivery, as described in detail above. Cells were subjected to differentiation assays as described above and in Example 1 in detail.

Embodiment 3

[0169] In vivo transplantation

[0170] The in vivo implantation of the cells of the present invention is useful for therapeutic applications in animals as well as for the treatment of human diseases such as type 1 diabetes caused by dysfunction of endocrine pancreatic tissue. Existing rodent models for these applications include insulin-dependent non-obese diabetic (NOD) mice, and mice or rats rendered diabetic by destruction of islets by treatment with streptozotocin (Lumelsky et al., 2001, Science 292 , 1389-1394; Soria et al., 2001, Diabetologia 44, 407-415). NOD mice have been used to engraft pancreatic islets and islets generated from pancreatic ductal stem cells (Soria et al., 2001, Diabetologia 44, 407-415; Lumelsky et al., 2001, Science 292, 1389-1394; Stegall et al., 2001). Differentiated cells of the invention expressing at least one genotypic or phenotypic characteristic of pancreatic cells are used to implant into NOD animals, which typically must be maintained o...

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Abstract

The invention provides cells, compositions and methods based on the differentiation of adipose tissue-derived stromal cells into a cell expressing at least one genotypic or phenotypic characteristic of a pancreas cell. The cells produced in the method are useful in providing a source of differentiated and functional cells for research, implantation, transplantation and development of tissue engineered products for the treatment of diseases of the pancreas and pancreatic tissue repair.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to US Serial No. 60 / 344,913, filed November 9, 2001. field of invention [0003] The present invention provides isolated adipose tissue-derived stromal cells induced to express at least one characteristic of pancreatic cells. Also provided are methods for treating pancreatic endocrine disorders. Background of the invention [0004] The endocrine cell mass of the islets includes four cell types classified according to their predominantly regulated secretory products. These cells include glucagon-producing alpha cells, insulin-producing beta cells, pancreatic polypeptide-producing gamma cells, and somatostatin-producing delta cells (Henquin, 2000, Diabetes 49, 1751-1760; Slack, 1995, Development 121 , 1569-1580). During development, these distinct cell populations are thought to arise from a common stem cell precursor associated with the pancreatic ductal epithelium (Rao et al., 1989, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/02A61K35/12A61K48/00A61L27/00A61P3/06A61P3/10A61P5/48C12N5/00C12N5/071C12N5/10
CPCC12N2510/00C12N5/0676C12N2501/115A61K2035/126A61K35/12C12N2506/1384C12N2502/1305A61P1/18A61P17/02A61P3/06A61P5/48A61P5/50A61P3/10C12N5/0652A61K35/39A01N1/02
Inventor B·奇塔姆Y-D·C·霍尔沃森J·M·金贝尔
Owner ARTECEL SCIENCE INC
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