Citation Details

Nucleofection is an efficient nonviral transfection technique for human bone marrow-derived mesenchymal stem cells

Authors	Aluigi M, Fogli M, Curti A, Isidori A, Gruppioni E, Chiodoni C, Colombo MP, Versura P, D'Errico-Grigioni A, Ferri E, Baccarani M, Lemoli RM
In	Stem Cells (2006) 24(2): 454-61
Cells used in publication	Mesench. stem (MSC), human    Human mesenchymal stem cells (MSC)    Stem Cells    Primary Cells    Species: human    Tissue Origin: bone marrow
Substrate	Plasmid (general)
Topics	Comparison to other transfection methods Differentiation of nucleofected cells Microscopy of nucleofected cells
Research Areas	Cancer Research/Cell Biology Dermatology/Tissue Engineering Immunology / Hematology
Relevant Products	Nucleofector Device Human MSC (Mesenchymal Stem Cell) Nucleofector Kit
Reporter Gene	GFP unspecified

Research Field

The authors have compared nucleofection and other non-viral methods for transfection of human mesenchymal stem cells.

Nucleofection Experiments

Using a green fluorescent protein reporter vector, the authors demonstrated a high transgene expression level using U-23 and C-17 pulsing programs: 73.7+/-2.9% and 42.5+/-3.4%, respectively. Cell recoveries and viabilities were 38.7+/-2.9%, 44.5+/-3.9% and 91.4+/-1.3%, 94.31+/-0.9% for U-23 and C-17, respectively. Overall, the transfection efficiencies were 27.4+/-2.9% (U-23) and 16.6+/-1.4% (C-17) as compared with 3.6+/-2.4% and 5.4+/-3.4% of other non-viral transfection systems such as FUGENE6 and DOTAP, respectively (p<0.005 for all comparisons). Nucleofection did not affect the immunophenotype of hMSCs, their normal differentiation potential and ability to inhibit T-cell alloreactivity. Moreover, the interleukin-12 gene could be successfully transfected into hMSC and the immunomodulatory cytokine was produced in great amount for at least 3 weeks without impairment of its biological activity.

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