title: High-efficiency lentiviral transduction of primary human CD34⁺ hematopoietic cells with low-dose viral inocula.
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.
authors.affiliation: Department of Life Sciences, Korea University, Seoul, 136-701, Korea, sangwon.park@childrens.harvard.edu.
authors.forename: Sang Won
authors.initials: SW
authors.lastname: Park
authors.forename: Chul-Woong
authors.initials: CW
authors.lastname: Pyo
authors.forename: Sang-Yun
authors.initials: SY
authors.lastname: Choi
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000235:genetics*; D004847:Epithelial Cells; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics*; D057809:HEK293 Cells; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism / Q000821:virology*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D041681:NIH 3T3 Cells; D013696:Temperature; D014161:Transduction, Genetic / Q000379:methods*
chemical_list: D018952:Antigens, CD34