Document Detail


Controlling mesenchymal stem cell gene expression using polymer-mediated delivery of siRNA.
MedLine Citation:
PMID:  23020123     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
siRNA treatment has great promise to specifically control gene expression and select cell behaviors but has delivery challenges limiting its use. Particularly for applications in regenerative medicine, uniform and consistent delivery of siRNA to control gene expression and subsequent stem cell functions, such as differentiation, is paramount. Therefore, a diblock copolymer was examined for its ability to effectively deliver siRNA to mesenchymal stem cells (MSCs). The diblock copolymers, which are composed of cationic blocks for siRNA complexation, protection, and uptake and pH-responsive blocks for endosomal escape, were shown to facilitate nearly 100% MSC uptake of siRNA. This is vastly superior to a commercially available control, DharmaFECT, which resulted in only ~60% siRNA positive MSCs. Moreover, the diblock copolymer, at conditions that result in excellent knockdown (down to ~10% of control gene expression), was cytocompatible, causing no negative effects on MSC survivability. In contrast, DharmaFECT/siRNA treatment resulted in only ~60% survivability of MSCs. Longitudinal knockdown after siRNA treatment was examined and protein knockdown persists for ~6 days regardless of delivery system (diblock copolymer or DharmaFECT). Finally, MSC phenotype and differentiation capacity was examined after treatment with control siRNA. There was no statistically significant differences on cell surface markers of diblock copolymer/siRNA or DharmaFECT/siRNA-treated or cells measured 2 weeks after siRNA delivery compared to untreated cells. Upon differentiation with typical media/culture conditions to adipogenic, chondrogenic, and osteogenic lineages and examination of histological staining markers, there was no discernible differences between treated and untreated cells, regardless of delivery mechanism. Thus, diblock copolymers examined herein facilitated uniform siRNA treatment of MSCs, inducing siRNA-specific gene and protein knockdown without adversely affecting MSC survival or differentiation capacity and therefore show great promise for use within regenerative medicine applications.
Authors:
Danielle S W Benoit; Molly E Boutin
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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:  2012-10-11
Journal Detail:
Title:  Biomacromolecules     Volume:  13     ISSN:  1526-4602     ISO Abbreviation:  Biomacromolecules     Publication Date:  2012 Nov 
Date Detail:
Created Date:  2012-11-12     Completed Date:  2013-06-05     Revised Date:  2014-04-02    
Medline Journal Info:
Nlm Unique ID:  100892849     Medline TA:  Biomacromolecules     Country:  United States    
Other Details:
Languages:  eng     Pagination:  3841-9     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Adipogenesis / genetics
Bone Marrow Cells / cytology*
Cell Differentiation / genetics
Cell Lineage
Chondrogenesis / genetics
Gene Expression
Gene Expression Regulation
Humans
Mesenchymal Stromal Cells / cytology*
Osteogenesis / genetics
Polymers / metabolism
RNA Interference
RNA Transport*
RNA, Small Interfering / genetics*
Grant Support
ID/Acronym/Agency:
P40 OD011050/OD/NIH HHS; P40 RR017447/RR/NCRR NIH HHS; P40RR017447/RR/NCRR NIH HHS
Chemical
Reg. No./Substance:
0/Polymers; 0/RNA, Small Interfering
Comments/Corrections

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


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