Document Detail


A bilayer construct controls adipose-derived stem cell differentiation into endothelial cells and pericytes without growth factor stimulation.
MedLine Citation:
PMID:  21083419     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
This work describes the differentiation of adipose-derived mesenchymal stem cells (ASC) in a composite hydrogel for use as a vascularized dermal matrix. Our intent is that such a construct could be utilized following large-surface-area burn wounds that require extensive skin grafting and that are limited by the availability of uninjured sites. To develop engineered skin replacement constructs, we are pursuing the use of ASC. We have established that a PEGylated fibrin gel can provide a suitable environment for the proliferation of ASC over a 7 day time course. Further, we have demonstrated that PEGylated fibrin can be used to control ASC differentiation toward vascular cell types, including cells characteristic of both endothelial cells and pericytes. Gene expression analysis revealed strong upregulation of endothelial markers, CD31, and von Willebrand factor, up to day 11 in culture with corresponding evidence of protein expression demonstrated by immunocytochemical staining. ASC were not only shown to express endothelial cell phenotype, but a subset of the ASC expressed pericyte markers. The NG2 gene was upregulated over 11 days with corresponding evidence for the cell surface marker. Platelet-derived growth factor receptor beta gene expression decreased as the multipotent ASC differentiated up to day 7. Increased receptor expression at day 11 was likely due to the enhanced pericyte gene expression profile, including increased NG2 expression. We have also demonstrated that when cells are loaded onto chitosan microspheres and sandwiched between the PEGylated fibrin gel and a type I collagen gel, the cells can migrate and proliferate within the two different gel types. The matrix composition dictates the lineage specification and is not driven by soluble factors. Utilizing an insoluble bilayer matrix to direct ASC differentiation will allow for the development of both vasculature as well as dermal connective tissue from a single population of ASC. This work underscores the importance of the extracellular matrix in controlling stem cell phenotype. It is our goal to develop layered composites as wound dressings or vascularized dermal equivalents that are not limited by nutrient diffusion.
Authors:
Shanmugasundaram Natesan; Ge Zhang; David G Baer; Thomas J Walters; Robert J Christy; Laura J Suggs
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.     Date:  2011-01-04
Journal Detail:
Title:  Tissue engineering. Part A     Volume:  17     ISSN:  1937-335X     ISO Abbreviation:  Tissue Eng Part A     Publication Date:  2011 Apr 
Date Detail:
Created Date:  2011-03-23     Completed Date:  2011-08-03     Revised Date:  2012-09-24    
Medline Journal Info:
Nlm Unique ID:  101466659     Medline TA:  Tissue Eng Part A     Country:  United States    
Other Details:
Languages:  eng     Pagination:  941-53     Citation Subset:  IM    
Affiliation:
Regenerative Medicine Research Program, US Army Institute of Surgical Research, Fort Sam, Houston, Texas, USA.
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MeSH Terms
Descriptor/Qualifier:
Adipose Tissue / cytology*
Animals
Cell Differentiation / physiology
Cells, Cultured
Endothelial Cells / cytology*,  ultrastructure
Fibrin / chemistry
Immunohistochemistry
Microscopy, Electron, Transmission
Pericytes / cytology*,  ultrastructure
Polyethylene Glycols / chemistry
Polymerase Chain Reaction
Rats
Stem Cells / cytology*,  ultrastructure
Tissue Engineering
Grant Support
ID/Acronym/Agency:
P01AG19316/AG/NIA NIH HHS; P30 AG013319/AG/NIA NIH HHS; P30 CA54174/CA/NCI NIH HHS
Chemical
Reg. No./Substance:
0/Polyethylene Glycols; 9001-31-4/Fibrin
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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