Optimization of a gene electrotransfer method for mesenchymal stem cell transfection

Area of Science:

• Biotechnology
• Molecular Biology
• Cell Biology

Background:

• Gene electrotransfer is a nonviral method for transient gene expression.
• Mesenchymal stem cells (MSCs) are multipotent cells with therapeutic potential.
• Optimizing gene electrotransfer into MSCs is crucial for their application.

Purpose of the Study:

• To establish optimal conditions for reporter gene electrotransfer into rat bone marrow-derived MSCs.
• To assess the impact of electrotransfer on MSC viability and multipotency.
• To evaluate the duration of transgene expression in MSCs.

Main Methods:

• Optimization of electric pulse parameters (intensity, type, buffer conductivity, temperature).
• Electrotransfer of the lacZ reporter gene into MSCs.
• Assessment of beta-galactosidase expression and MSC differentiation potential.

Main Results:

• Optimal lacZ electrotransfer into MSCs achieved at 1500 V cm(-1) with pre-incubation in Spinner's MEM at 22°C.
• Beta-galactosidase expression observed in 29+/-3% of cells 48 hours post-transfection.
• Electroporation did not impair MSC differentiation into osteoblastic, adipogenic, or chondrogenic lineages.

Conclusions:

• Efficient and reproducible transgene electrotransfer into MSCs is feasible.
• The established conditions preserve MSC viability and multipotency.
• Gene electrotransfer is a promising technique for MSC-based gene therapy.