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Updated: Aug 16, 2026
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Isolation, Expansion, and Nucleofection of Neural Stem Cells from Adult Murine Subventricular Zone
Published on: June 14, 2024
Nucleofection is an efficient nonviral transfection technique for human bone marrow-derived mesenchymal stem cells
Michela Aluigi1, Miriam Fogli, Antonio Curti
1Institute of Hematology and Medical Oncology L. e A. Seràgnoli, Via Massarenti, 9, 40137 Bologna, Italy. maluigi@biocfarm.unibo.it
Stem Cells (Dayton, Ohio)
|August 16, 2005
Summary
Nucleofection, a nonviral gene transfer method, efficiently engineers human mesenchymal stem cells (hMSCs). This technique offers a safer alternative to viral vectors for stem cell modification and therapeutic applications.
Area of Science:
- Cell Biology
- Biotechnology
- Gene Therapy
Background:
- Viral vectors are efficient for gene delivery into stem cells but pose safety risks.
- Nonviral methods are generally inefficient for primary cells, including adult stem cells.
- Nucleofector technology is a nonviral electroporation method known for transfecting difficult cell types.
Purpose of the Study:
- To evaluate the efficacy of Nucleofector technology for engineering human bone marrow-derived mesenchymal stem cells (hMSCs).
- To compare Nucleofection with other nonviral transfection systems for hMSCs.
- To assess the impact of Nucleofection on hMSC characteristics and functionality.
Main Methods:
- Application of Nucleofector technology using U-23 and C-17 pulsing programs for hMSC transfection.
- Utilized a green fluorescent protein (GFP) reporter vector to quantify transgene expression.
- Assessed cell recovery, viability, immunophenotype, differentiation potential, and T-cell inhibition capacity.
- Transfected the interleukin-12 (IL-12) gene into hMSCs to evaluate functional protein production.
Main Results:
- High transgene expression levels (73.7% with U-23, 42.5% with C-17) were achieved in hMSCs.
- Nucleofection demonstrated significantly higher transfection efficiencies (27.4% with U-23, 16.6% with C-17) compared to FUGENE6 and DOTAP.
- Nucleofection did not adversely affect hMSC immunophenotype, differentiation potential, or immunomodulatory functions.
- Successfully transfected IL-12 gene into hMSCs, resulting in sustained production of biologically active cytokine for over 3 weeks.
Conclusions:
- Nucleofection is a highly efficient and safe nonviral transfection technique for human mesenchymal stem cells.
- Engineered hMSCs can serve as effective cellular vehicles for delivering therapeutic biological agents.
- This method provides a promising alternative for stem cell-based gene therapy and regenerative medicine.
