Enhanced lipoplex‐mediated gene expression in mesenchymal stem cells using reiterated nuclear localization sequence peptides

Hoare, Melissa; Greiser, Udo; Schu, Sabine; Mashayekhi, Kaveh; Aydogan, Emrah; Murphy, Mary; Barry, Frank; Ritter, Thomas; O’Brien, Timothy

doi:10.1002/jgm.1426

Cited by 41 publications

(44 citation statements)

References 25 publications

(42 reference statements)

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“…Most of the studies focusing on the transfection of animal cells in general and human MSC in particular have been using initial cell densities higher than 7500 cells/cm 2 mainly in order to maximize the cell numbers available for analysis (e.g., flow cytometry) (Gheisari et al, 2008;Hoare et al, 2010;Madeira et al, 2010). These plating densities are higher than those commonly used in MSC expansion studies.…”

mentioning

confidence: 87%

“…In an effort to minimize surface area limitation to the cultured cells and to ensure that the majority of the cells were actively dividing at the time of transfection, MSC were plated at lower cell densities compared to other transfection studies using human MSC (Gheisari et al, 2008;Hoare et al, 2010;Madeira et al, 2010) and lipofected 72 hr upon seeding.…”

Section: Efficiency Of Liposome-mediated Gene Transfer To Msc From DImentioning

confidence: 99%

“…To the best of our knowledge, this represents the first systematic study of the gene delivery performance of a nonviral vector to MSC of different human sources, which are relevant for cell/gene therapy settings. By plating cells at a lower initial cell density compared to previous studies in literature (Gheisari et al, 2008;Hoare et al, 2010;Madeira et al, 2010), a key step for the successful long-term cultivation of human MSC (Sekiya et al, 2002), lipofection efficiencies were maximized. Cell survival, cellular division kinetics, differentiation, and clonogenic potential, as well as the immunophenotype of the engineered human MSC, were also studied.…”

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confidence: 99%

“…Though the mechanism of gene delivery inside the cells is not completely understood, it is accepted that this method is dependent on both cell division and endocytosis rate (Gresch et al, 2004). Regarding human MSC, transfection using the lipofection method has led to moderate levels of transgenepositive cells (30-40%) (Hoare et al, 2010). Higher dosage of cationic agents generally increased transfection efficacy but were also associated with low cell survival and toxicity (McMahon et al, 2006;Clements et al, 2007;Farrell et al, 2007;Gheisari et al, 2008).…”

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confidence: 99%

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Direct Head-To-Head Comparison of Cationic Liposome-Mediated Gene Delivery to Mesenchymal Stem/Stromal Cells of Different Human Sources: A Comprehensive Study

Boura

Santos

Gimble

et al. 2013

Human Gene Therapy Methods

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Nonviral gene delivery to human mesenchymal stem/stromal cells (MSC) can be considered a very promising strategy to improve their intrinsic features, amplifying the therapeutic potential of these cells for clinical applications. In this work, we performed a comprehensive comparison of liposome-mediated gene transfer efficiencies to MSC derived from different human sources-bone marrow (BM MSC), adipose tissue-derived cells (ASC), and umbilical cord matrix (UCM MSC). The results obtained using a green fluorescent protein (GFP)-encoding plasmid indicated that MSC isolated from BM and UCM are more amenable to genetic modification when compared to ASC as they exhibited superior levels of viable, GFP + cells 48 hr post-transfection, 58 -7.1% and 54 -3.8%, respectively, versus 33 -4.7%. For all cell sources, high cell recoveries (&50%) and viabilities (> 85%) were achieved, and the transgene expression was maintained for 10 days. Levels of plasmid DNA uptake, as well as kinetics of transgene expression and cellular division, were also determined. Importantly, modified cells were found to retain their characteristic immunophenotypic profile and multilineage differentiation capacity. By using the lipofection protocol optimized herein, we were able to maximize transfection efficiencies to human MSC (maximum of 74% total GFP + cells) and show that lipofection is a promising transfection strategy for MSC genetic modification, especially when a transient expression of a therapeutic gene is required. Importantly, we also clearly demonstrated that intrinsic features of MSC from different sources should be taken into consideration when developing and optimizing strategies for MSC engineering with a therapeutic gene.

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mentioning

confidence: 87%

Section: Efficiency Of Liposome-mediated Gene Transfer To Msc From DImentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Direct Head-To-Head Comparison of Cationic Liposome-Mediated Gene Delivery to Mesenchymal Stem/Stromal Cells of Different Human Sources: A Comprehensive Study

Boura

Santos

Gimble

et al. 2013

Human Gene Therapy Methods

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show abstract

“…Transferin-conjugated lipoplexes were also used to evaluate specific gene delivery to a metastatic mammary carcinoma cell line, although the results did not demonstrate a significant increase in cytotoxicity with modified lipoplexes in comparison with nonmodified lipoplex (Lopez-Barcons et al, 2005). Another strategy based on ex-vivo gene therapy assessed the challenge of nuclear entry by using plasmids transfected with a cationic lipid vector in MSC (Hoare et al, 2010). To enhance gene expression in MSC, they used NLS peptides to direct the plasmid to the nucleus.…”

Section: Lipoplexesmentioning

confidence: 99%