Abstract
Lentivirus-based vectors have the potential to transduce non-dividing primary stem cells. However, primary cells tend to be less susceptible to manipulation and require a high concentration of virus inoculum. Furthermore, increasing the concentration of the lentivirus inoculum may raise safety risks. Therefore, to develop a technique that allows high transduction efficiency at low multiplicities of infection (MOIs), we optimized a lentivirus-based system for cell lines and primary cells by determining the best condition using various parameters. When progenitor cell assays were conducted using human CD34+ bone marrow and mononuclear cells, the transduction condition yielded a great number of eGFP+ colonies with lower-dose viral inocula compared to that of current lentivirus-based transduction technologies. In conclusion, this system is anticipated to produce stable expression of a gene introduced into primary cells for preclinical studies with lower safety risks.
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Acknowledgments
This work was supported by the Korea Research Foundation Grant (KRF08-C00557) and a Grant of the Korea Healthcare Technology R&D Project of the Ministry for Health and Welfare, Republic of Korea (A091007).
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Park, S.W., Pyo, CW. & Choi, SY. High-efficiency lentiviral transduction of primary human CD34+ hematopoietic cells with low-dose viral inocula. Biotechnol Lett 37, 281–288 (2015). https://doi.org/10.1007/s10529-014-1678-z
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DOI: https://doi.org/10.1007/s10529-014-1678-z