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Multipotent mesenchymal stromal cells: optimization and comparison of five cationic polymer-based gene delivery
Y Gheisari1, M Soleimani, K Azadmanesh
1Department of Molecular Medicine, Pasteur Institute of Iran, Tehran, Iran.
Cytotherapy
|December 6, 2008
Summary
Optimizing gene delivery to multipotent mesenchymal stromal cells (MSC) is crucial for regenerative medicine. Lipofectamine 2000 and Polyfect demonstrated moderate transfection rates and acceptable cell viability in rat bone marrow-derived MSC.
Area of Science:
- Regenerative Medicine
- Cell Biology
- Gene Therapy
Background:
- Multipotent mesenchymal stromal cells (MSC) are key for regenerative medicine.
- MSC can be genetically modified or used as gene delivery vehicles.
- Effective gene delivery methods are essential for MSC applications.
Purpose of the Study:
- To systematically optimize five cationic polymer-based gene delivery methods for rat bone marrow-derived MSC.
- To evaluate transfection yield and cell viability for each method.
Main Methods:
- Rat bone marrow MSC isolation and characterization (differentiation, surface markers).
- Optimization of five gene delivery reagents: Lipofectamine 2000, Effecten, Superfect, Polyfect, and FuGENE HD.
- Measurement of transfection rates and cell viability at various reagent ratios and amounts.
Main Results:
- Isolated MSC confirmed as authentic (osteoblasts, adipocytes, chondroblasts; CD90+, CD73+, CD31-, CD45-, CD11b-, VEGFR2-).
- Optimal transfection rates: Lipofectamine 2000 (19.60%), Polyfect (16.29%), Superfect (9.59%), Effecten (8.72%), FuGENE HD (5.18%).
- All reagents showed <20% toxicity.
Conclusions:
- Lipofectamine 2000 and Polyfect offer moderate transfection and acceptable viability for MSC gene delivery.
- Further improvement in transfection efficiency is possible via FSC-SSC gating for live cell sorting.
