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KMID : 0545120050150020451
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Journal of Microbiology and Biotechnology
2005 Volume.15 No. 2 p.451 ~ p.454
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Highly Efficient Gene Delivery into Transfection-Refractory Neuronal and Astroglial Cells Using a Retrovirus-Based Vector
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Kim BO
Pyo SN
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Abstract
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Introduction of foreign genes into brain cells such as neurons and astrocytes is a powerful approach to study the gene function and regulation in the neuroscience field. Calcium phosphate precipitates have been shown to cause cytotoxicity in some mammalian cells and brain cells thus leading to low transfection efficiency. Here we describe a retrovirus-mediated gene delivery method to transduce foreign genes into brain cells. In an attempt to achieve higher gene delivery efficiency in these cells we made several changes to the original method including (1) use of a new packaging cell line Phoenix ampho cells (2) transfection of pMX retroviral DNA (3) inclusion of 25 mM chloroquine in the transduction and (4) 3- 5 h incubation of retroviruses with target cells. The results showed that the modified protocol resulted in a range of 40- 60% gene delivery efficiency in neurons and astrocytes. Furthermore these results suggest the potential of the retrovirus-mediated gene delivery protocol being modified and adapted for other transfection-refractory cell lines and primary cells.
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