Park Sang Won, Pyo Chul-Woong, Choi Sang-Yun
Department of Life Sciences, Korea University, Seoul, 136-701, Korea,
Biotechnol Lett. 2015 Feb;37(2):281-8. doi: 10.1007/s10529-014-1678-z. Epub 2014 Sep 26.
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.
基于慢病毒的载体有转导非分裂原代干细胞的潜力。然而，原代细胞往往对操作不太敏感，并且需要高浓度的病毒接种物。此外，增加慢病毒接种物的浓度可能会增加安全风险。因此，为了开发一种在低感染复数（MOI）下具有高转导效率的技术，我们通过使用各种参数确定最佳条件，对用于细胞系和原代细胞的基于慢病毒的系统进行了优化。当使用人CD34(+)骨髓和单核细胞进行祖细胞分析时，与当前基于慢病毒的转导技术相比，该转导条件在较低剂量病毒接种物的情况下产生了大量eGFP(+)集落。总之，该系统有望在安全性风险较低的情况下，使导入原代细胞的基因在临床前研究中实现稳定表达。
