Direct head-to-head comparison of cationic liposome-mediated gene delivery to mesenchymal stem/stromal cells of different human sources: a comprehensive study.
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.
Department of Bioengineering and IBB-Institute for Biotechnology and Bioengineering, Instituto Superior Técnico, Technical University of Lisbon, Lisboa 1049-001, Portugal.
Joana S
JS
Boura
Francisco Dos
FD
Santos
Jeffrey M
JM
Gimble
Carla M P
CM
Cardoso
Catarina
C
Madeira
Joaquim M S
JM
Cabral
Cláudia Lobato da
CL
Silva
D017667:Adipocytes / Q000166:cytology / Q000378:metabolism; D000328:Adult; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism; D002412:Cations; D002454:Cell Differentiation; D049109:Cell Proliferation; D056915:DNA Copy Number Variations; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D008081:Liposomes; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D008875:Middle Aged; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D019076:Transgenes; D014470:Umbilical Cord / Q000166:cytology / Q000378:metabolism
D002412:Cations; D008081:Liposomes; D049452:Green Fluorescent Proteins