Nonviral Gene Delivery to Mesenchymal Stem Cells Using Cationic Liposomes for Gene and Cell Therapy

Madeira, Catarina; Mendes, Rui D.; Ribeiro, Sofia; Boura, Joana S.; Aires-Barros, M. Raquel; Silva, C. L. da; Cabral, Joaquim M. S.

doi:10.1155/2010/735349

Cited by 79 publications

(78 citation statements)

References 37 publications

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“…Under determined conditions, we observed that the ratio of cell viability and transfection in both analyzed stem cell types were inverse, as has been reported by other groups (Madeira et al 2010;Gheisari et al 2008). The largest number of viable hMSCs was found after transfection with HiPerFect or Oligofectamine with nearly the lowest transfection efficiencies.…”

Section: Discussionsupporting

confidence: 85%

A comparative study on nonviral genetic modifications in cord blood and bone marrow mesenchymal stem cells

et al. 2012

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The focus of both clinical and basic studies on stem cells is increasing due to their potentials in regenerative medicine and cell-based therapies. Recently stem cells have been genetically modified to enhance an existing character in or to bring a new property to them. However, accomplishment of declared goals requires detailed knowledge about their molecular characteristics which could be achieved by genetic modifications mostly through nonviral transfection strategies. Capable of differentiating into multiple cells, human unrestricted somatic stem cells (hUSSCs) and human mesenchymal stem cells (hMSCs) seem to be suitable candidates for transfection approaches. Involvement of microRNAs (miRNAs) in many biological processes makes their transfection evaluation valuable. Herein we investigated the efficacy and toxicity of four typically used transfection reagents (Arrest-In, Lipofectamine 2000, Oligofectamine and HiPerfect) systematically to deliver fluorescent labeled-miRNA and Green Fluorescent Protein (GFP) expressing plasmid into hUSSCs and hMSCs. The authenticity of stem cells was verified by differentiation experiments along with flow cytometry of surface markers. Our study revealed that stemness properties of these stem cells were not affected by transient transfection. Moreover the ratios of cell viability and transfection efficiency in both analyzed stem cells were reversed. Considering cell viability, the highest fraction of GFP-expressing cells was obtained using Oligofectamine (*50%) while the highest transfection rate of miRNA was achieved by Lipofectamine 2000 (*90%). Moreover dependency of hMSCs to size of transfected nucleic acid and timedependency of Oligofectamine and their affection on the yield of transfection were observed. Cytotoxicity assessments also showed that hUSSCs are sensitive to HiPerFect. In addition cells treated by Lipofectamine showed morphological changes. Representing the efficient nucleic acid transfection, our research facilitates comprehensive genetic modification of stem cells and demonstrates powerful approaches to understand stem cell molecular regulation mechanisms, Electronic supplementary material The online version of this article

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Section: Discussionsupporting

confidence: 85%

A comparative study on nonviral genetic modifications in cord blood and bone marrow mesenchymal stem cells

et al. 2012

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“…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). Recently, our group reported promising results using this reagent for gene delivery to BM MSC (Madeira et al, 2010).…”

mentioning

confidence: 97%

“…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%

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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“…Next, DNA construct was mixed with various lipid components including cationic, neutral lipids. Lipid based systems are exceedingly expected to be very compatible with cellular membranes, owing to the compositional features replicating the plasma membrane (24). However, it was evident that the system was deficient in active targeting once mixed with variable sizes of DNA molecules.…”

Section: Chemical (Polymer Based) Methodsmentioning

confidence: 99%

Delivering Factors for Reprogramming a Somatic Cell to Pluripotency

2012

Int J Stem Cells

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An adult cell originates from stem cell. The stem cell is usually categorized into three species including an embryonic stem cell (ESc), an adult stem cell, and an induced stem cell (iPSc). iPSc features pluripotency, which is meant to be differentiated into any types of cells. Accordingly, it is much attractive to anyone who pursuit a regenerative medicine, owing to the potential almighty. They are simply produced by reprogramming a somatic cell via a transfer of transcription factors. The efficiency and productivity of iPS are considerably subject to delivering methods of exogenous genes into a variety of targeted mammalians. Conventional and well-run gene delivery techniques have been reviewed here. This details the methods and principles of delivery factors and provides an overview of the research, with an emphasis on their potential for use as clinical therapeutic platforms.

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Nonviral Gene Delivery to Mesenchymal Stem Cells Using Cationic Liposomes for Gene and Cell Therapy

Cited by 79 publications

References 37 publications

A comparative study on nonviral genetic modifications in cord blood and bone marrow mesenchymal stem cells

A comparative study on nonviral genetic modifications in cord blood and bone marrow mesenchymal stem cells

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

Delivering Factors for Reprogramming a Somatic Cell to Pluripotency

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