Gene delivery to human bone marrow mesenchymal stem cells by microporation

Madeira, Catarina; Ribeiro, Sofia; Pinheiro, Irina; Martins, Sofia Aires M.; Andrade, Pedro de; Silva, Cláudia L. da

doi:10.1016/j.jbiotec.2010.11.002

Cited by 34 publications

(27 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The metabolic burden imposed by the presence of large amounts of plasmid inside the cells within this period may be the main reason for this cell division arrest. Similar results were obtained by Madeira and colleagues after BM MSC microporation (Madeira et al, 2011). On the other hand, as control cells rapidly reach high levels of confluence, cell division is stopped due to surface area limitations approximately two generations before GFP + cells.…”

supporting

confidence: 86%

“…Remarkably, with this strategy, we were able to increase by two-to three-fold liposome-mediated gene delivery efficiencies to human MSC when compared to previous studies, where cells are seeded at subconfluent levels (80-90%) and transfected the day after (Hoare et al, 2010;Madeira et al, 2010). In fact, the levels of gene expression obtained are comparable to the ones reported with nucleofection (approximately 70%) (Aluigi et al, 2006;Aslan et al, 2006;Flanagan et al, 2012) and microporation (60-80%) (Wang et al, 2008;Madeira et al, 2011).…”

supporting

confidence: 70%

“…The percentage of viable cells was estimated using the trypan blue dye exclusion method. Cell recovery and yield of transfection were determined using equations previously described (Madeira et al, 2011).…”

Section: Liposome-mediated Transfectionmentioning

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

Self Cite

View full text Add to dashboard Cite

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.

show abstract

supporting

confidence: 86%

supporting

confidence: 70%

See 1 more Smart Citation

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

Self Cite

View full text Add to dashboard Cite

show abstract

“…This transfection technique does not affect the proliferative, differentiative or migratory activity of the cells. 34 …”

Section: Cell Cultures and Transfectionmentioning

confidence: 99%

Down-regulation of 21A Alu RNA as a tool to boost proliferation maintaining the tissue regeneration potential of progenitor cells

et al. 2016

View full text Add to dashboard Cite

ABSTRACT21A is an Alu non-coding (nc) RNA transcribed by RNA polymerase (pol) III. While investigating the biological role of 21A ncRNA we documented an inverse correlation between its expression level and the rate of cell proliferation. The downregulation of this ncRNA not only caused a boost in cell proliferation, but was also associated to a transient cell dedifferentiation, suggesting a possible involvement of this RNA in cell dedifferentiation/reprogramming. In this study, we explored the possibility to enhance proliferation and dedifferentiation of cells of interest, by 21A down-regulation, using a mixture of chemically modified Anti-21A RNAs. Our results confirmed the validity of this approach that allows the amplification of specific cell populations, in a controlled manner and without inducing permanent effects. In addition to induce cell proliferation, the procedure did not decrease the tissue regeneration potential of progenitor cells in two different cell systems.

show abstract

“…Even in the field of viral vectors, attempts have been made to combine their application with immunomodulating agents (Ikeda et al, 2000), still obvious problems exist with this approach (Zhou et al, 2004). Similarly, in the area of non-viral gene transfer, researchers are generating new transfection methods/strategies with a number of new products becoming commercially available every year (Donkuru et al, 2010;Haag et al, 2009;Madeira et al, 2011). Yet, an ideal agent that satisfies the requirements for application with the relevant target cells for OA gene therapy is not in sight.…”

Section: Obstacles and Challenges In Clinical Application Of Gene Thementioning

confidence: 99%

Conditioning and Scaffolding of Chondrocytes: Smart Steps Towards Osteoarthritis Gene Therapy

Farooq¹,

Lang²,

Sieber³

et al. 2011

Gene Therapy Applications

View full text Add to dashboard Cite

Gene delivery to human bone marrow mesenchymal stem cells by microporation

Cited by 34 publications

References 35 publications

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

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

Down-regulation of 21A Alu RNA as a tool to boost proliferation maintaining the tissue regeneration potential of progenitor cells

Conditioning and Scaffolding of Chondrocytes: Smart Steps Towards Osteoarthritis Gene Therapy

Contact Info

Product

Resources

About