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Updated: Sep 12, 2026
Production of Replication-Defective Retrovirus by Transient Transfection of 293T cells
Published on: December 4, 2007
Human serum-resistant retroviral vector particles from galactosyl (alpha1-3) galactosyl containing nonprimate cell
J M Mason1, D E Guzowski, L O Goodwin
1Viral Vector Laboratory, Department of Research, North Shore University Hospital-New York University School of Medicine, Manhasset, NY 11030, USA.
Abstract:
Retroviral vector particles (RVP) which are resistant to inactivation by human serum will be needed for many in vivo gene therapy applications. Murine-based producer cell lines generate RVP which are inactivated by human serum, reportedly due to the presence of the galactosyl (alpha1-3) galactosyl carbohydrate moiety (alphaGal) on these and other nonprimate producer cells and RVP. Consequently, human cells (which lack the alphaGal moiety) have been developed as producer cell lines for generation of human serum-resistant RVP. In this study, we report that contrary to earlier reports, the presence of the alphaGal moiety on producer cells and RVP does not necessarily correlate with cell killing or RVP inactivation by human serum. We show that the alphaGal-positive ferret brain cell line, Mpf, is an excellent basal cell line for generation of RVP which have titers and serum resistance levels equal to or greater than RVP produced in human cell lines such as HT1080. Therefore, packaging cell lines need not be limited to those of human or primate origin for production of human serum-resistant RVP.
