Nonviral Gene Delivery to Neural Stem Cells with Minicircles by Microporation

The main purpose of this work was to evaluate the transfection of novel DNA vectors, minicircles (mC), on embryonic stem cell-derived neural stem cells (NSC). We demonstrated that by combining microporation with mC, 75% of NSC expressing a transgene is achieved without compromising cell survival, mo...

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Bibliographic Details
Published in:Biomacromolecules 2013-05, Vol.14 (5), p.1379-1387
Main Authors: Madeira, Catarina, Rodrigues, Carlos A. V, Reis, Mónica S. C, Ferreira, Filipa F. C. G, Correia, Raquel E. S. M, Diogo, Maria M, Cabral, Joaquim M. S
Format: Article
Language:English
Subjects:
Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy
Animals
Applied cell therapy and gene therapy
Biological and medical sciences
Cell Differentiation
Cell Line
Cell Proliferation
Cell Survival
DNA - genetics
DNA Copy Number Variations
Electroporation
Genetic Vectors
Green Fluorescent Proteins - genetics
Medical sciences
Mice
Neural Stem Cells - cytology
Neural Stem Cells - metabolism
Plasmids
Transfection - instrumentation
Transfection - methods
Transfusions. Complications. Transfusion reactions. Cell and gene therapy
Transgenes
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Summary:The main purpose of this work was to evaluate the transfection of novel DNA vectors, minicircles (mC), on embryonic stem cell-derived neural stem cells (NSC). We demonstrated that by combining microporation with mC, 75% of NSC expressing a transgene is achieved without compromising cell survival, morphology, and differentiation potential. When comparing mC with their plasmid DNA (pDNA) counterparts, both gave rise to similar transfection levels but cells harboring mC showed 10% higher cell viability, maintaining 90% of survival at least for 10 days. Long-term analysis showed that NSC harbor a higher number of mC copies and consequently exhibit higher transgene expression when compared to their pDNA counterpart. Taken together, our results offer the first insights on the use of mC as a novel and safe strategy to genetically engineer NSC envisaging their use as biopharmaceuticals in clinical settings for the treatment of neurodegenerative or neurological diseases.
ISSN:1525-7797
1526-4602
DOI:10.1021/bm400015b