Nonviral gene delivery to mesenchymal stem cells using cationic liposomes for gene and cell therapy.
Mesenchymal stem cells (MSCs) hold a great promise for application in several therapies due to their unique biological characteristics. In order to harness their full potential in cell-or gene-based therapies it might be advantageous to enhance some of their features through gene delivery strategies. Accordingly, we are interested in developing an efficient and safe methodology to genetically engineer human bone marrow MSC (BM MSC), enhancing their therapeutic efficacy in Regenerative Medicine. The plasmid DNA delivery was optimized using a cationic liposome-based reagent. Transfection efficiencies ranged from approximately 2% to approximately 35%, resulting from using a Lipid/DNA ratio of 1.25 with a transgene expression of 7 days. Importantly, the number of plasmid copies in different cell passages was quantified for the first time and approximately 20,000 plasmid copies/cell were obtained independently of cell passage. As transfected MSC have shown high viabilities (>90%) and recoveries (>52%) while maintaining their multipotency, this might be an advantageous transfection strategy when the goal is to express a therapeutic gene in a safe and transient way.
Madeira
C
C
IBB-Institute for Biotechnology and Bioengineering, Centre for Biological and Chemical Engineering, Instituto Superior Técnico, Av. Rovisco Pais, Lisboa, Portugal.
Mendes
R D
RD
Ribeiro
S C
SC
Boura
J S
JS
Aires-Barros
M R
MR
da Silva
C L
CL
Cabral
J M S
JM
D000328:Adult; D002412:Cations; D002470:Cell Survival; D003114:Colony-Forming Units Assay; D005434:Flow Cytometry; D015316:Genetic Therapy; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D016130:Immunophenotyping; D008081:Liposomes / Q000378:metabolism*; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D039902:Multipotent Stem Cells / Q000166:cytology / Q000378:metabolism; D014162:Transfection / Q000379:methods*; D014780:Viruses / Q000235:genetics
D002412:Cations; D008081:Liposomes; D049452:Green Fluorescent Proteins