Adipose stromal stem cells for tissue and vascular modification

Inactive Publication Date: 2005-11-10
TISSUE GENESIS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0029]FIG. 16 is a graph showing that on day 10, adipose stromal cell treated animals had a s

Problems solved by technology

Coronary artery disease (CAD) is a major cause of morbidity and mortality, requiring bypass surgery or angioplasty in almost 1,000,000 patients/year in the USA.
While some of these patients form collateral vessels as alternative pathways for blood supply, thus ameliorating or preventing ischemic myocardial damage, many do not form the vascular networks to sufficiently compensate for the loss of the original blood supply.
Although recently there has been some success in the reduction of co

Method used

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  • Adipose stromal stem cells for tissue and vascular modification
  • Adipose stromal stem cells for tissue and vascular modification
  • Adipose stromal stem cells for tissue and vascular modification

Examples

Experimental program
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Effect test

example 1

Isolation and Characterization of Stem Cells from Adipose Tissue

Isolation of Stem Cells From Adipose Tissue

[0116] To isolate adipose stromal cells (ASCs,) subcutaneous adipose tissue (5-15 grams) was obtained from obese patients undergoing a gastroplasty procedure. The tissue was suspended in PBS and dissected into, smaller pieces using a scalpel. Collagenase (Worthington Biochemical) was then added to the suspension and placed in a shaker at 37° C. for approximately 90 minutes. The digested sample was filtered through a 750 micron Nitex filter and a 50 micron filter, and rinsed with Dulbecco's Modified Eagle Media (DMEM) with 10% Fetal Bovine Serum (FBS). The filtered suspension was centrifuged at 200 g for 5 minutes and the top layer consisting of adipocytes was discarded. After a second spin at 300 g for 5 minutes, any remaining adipocytes in the top layer were again discarded. The cell pellet was re-suspended in red cell lysis buffer and centrifuged at 300 g after 5-10 minute...

example ii

Differentiation of Adipose Derived Stromal Cells

ASCs Can Develop a Vascular Phenotype

[0120] A major pathway by which ASCs can enhance angiogenesis is by differentiation of ASCs into a vascular cell phenotype. Since ASCs are already known to differentiate into a number of cell types like muscle, bone and neural cells, it was evaluated whether ASCs can develop phenotypes that correspond to either vascular endothelial cells or vascular smooth muscle cells. Plated adherent human ASCs were evaluated by flow cytometry for the expression of the stem cell marker CD34 and the endothelial marker VE-Cadherin, (FIG. 5). While the majority of cells are still CD34+ / VE-Cadherin, there has emerged a substantial proportion of cells that are CD34+ / VE-Cadherin+; this finding has been replicated consistently with samples from several donors. This is in contrast to freshly isolated ASCS, which express minimal VE-Cadherin (FIG. 3), prior to plating. The appearance of a CD34+ / VE-Cadherin+ cell populati...

example iii

Secretion of Growth Factors by Adipose Derived Stromal Cells

Secretion of Angiogenic Growth Factors by ASCs

[0124] Since stem and progenitor cells can secrete multiple growth factors (Majka M, et al. Blood. (2001)97:3075-85) and putative therapeutic utilities could in part be related to this endocrine function, the secretion of angiogenic growth factors by human subcutaneous ASCs was evaluated. Human subcutaneous adipose stromal cells were cultured in EGM-2-MV media to confluence, and then switched into growth-factor free basal media (EBM-2, Clonetics) for 72 hours. Cell supernatants were collected and subsequently assayed for angiogenic growth factors using ELISA and Multi-Analyte-Profiling kits (R&D Systems) for Vascular Endothelial Growth Factor (VEGF), Hepatocyte Growth Factor (HGF) and Granulocyte-Colony Stimulating Factor (G-CSF). The cell number was assessed at the end of the 72-hour period, and the secretion of growth factors is expressed in pg / 106 ASCs. The cell supernatan...

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Abstract

The present invention provides isolated adipose derived stromal cells and methods of use thereof.

Description

CROSS REFERENCE TO RELATED APPLICATION [0001] This application claims priority to U.S. provisional application 60 / 365,498 filed Mar. 19, 2002, the entire disclosure of which is incorporated by reference herein.FIELD OF THE INVENTION [0002] The present invention relates to the fields of cardiology, vascularization, and molecular biology. More specifically, methods are provided for isolating stem cells from adipose tissue, optionally inducing the stem cells to secrete endogenous or exogenously provided growth factors, and re-administering such stem cells to a patient for therapeutic benefit. BACKGROUND OF THE INVENTION [0003] Several publications and patent documents are cited throughout the specification in order to describe the state of the art to which this invention pertains. Each of these references is incorporated herein as though set forth in full. [0004] Coronary artery disease (CAD) is a major cause of morbidity and mortality, requiring bypass surgery or angioplasty in almost...

Claims

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Application Information

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IPC IPC(8): A61K35/12C12N5/0775
CPCA61K35/12C12N5/0653C12N5/0667A61P9/00C12N2506/1384C12N2506/08C12N5/0619C12N2501/105C12N2501/11C12N2501/115C12N2501/165C12N2506/13
Inventor MARCH, KEITH L.REHMAN, JALEES
Owner TISSUE GENESIS
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