Gene Delivery to Stem Cells by Combination of Chitosan-Graft-Polyethylenimine as a Gene Carrier and E-Cadherin-IgG Fcas an Extracellular Matrix
Abstract:The ability of embryonic stem cells and adult stem cells to differentiate into specific cell types holds immense potential for therapeutic use in cell and gene therapy. Realization of this potential depends on efficient protocols for genetic manipulation of stem cells. However, conventional ways of gene transfer to such progenitor cells suffer from a number of disadvantages particularly involving safety and efficacy issues. Here, we report on the safe and efficient non-viral gene carrier—chitosan-graft-polyethylenimine (CHI-g-PEI) and extracellular matrix—E-cadherin-IgG Fc-coated culture system. The cytotoxicity of CHI-g-PEI copolymer was evaluated, and the results indicated that CHI-g-PEI copolymer had low cytotoxicity than PEI 25 K as a control in F9 and EB3 embryonic stem cells. Also, at high N/P ratio, CHI-g-PEI/DNA complexes showed higher transfection efficiency than that of PEI 25 K. The transfection efficiency of EB3 embryonic stem cells cultured on the plate coated with a fusion protein of E-cadherin and IgG Fc domain to prevent formation of aggregated colonies, showed higher transfection efficiency than that of cultured on gelatin-coated plate, suggesting that combination of gene carrier and extracellular matrix will be an efficient way to develop of stem cell-based gene therapy.
Document Type: Research Article
Publication date: December 1, 2007
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