High-efficiency lentiviral transduction of primary human CD34+ hematopoietic cells with low-dose viral inocula
High-efficiency lentiviral transduction of primary human CD34+ hematopoietic cells with low-dose viral inocula
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低剂量病毒接种原代人 CD34+ 造血细胞的高效慢病毒转导
DOI：
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发表时间：
2014
影响因子：
2.7
通讯作者：
Sang‐Yun Choi
中科院分区：
文献类型：
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作者：
S. Park;Chul;Sang‐Yun Choi
基于慢病毒的载体具有使非分裂的原代干细胞增殖的潜力。然而，原代细胞倾向于不易受操作影响，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，为了开发在低感染复数（MOI）下允许高转导效率的技术，我们通过使用各种参数确定最佳条件来优化用于细胞系和原代细胞的基于慢病毒的系统。当使用人CD34+骨髓和单核细胞进行祖细胞测定时，与目前基于慢病毒的转导技术相比，转导条件产生了大量的eGFP+集落，具有较低剂量的病毒接种物。总之，预期该系统可稳定表达导入原代细胞的基因，用于临床前研究，且安全性风险较低。
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.
DOI：
10.1073/pnas.96.6.2988
发表时间：
1999-03-16
影响因子：
11.1
作者：
Case, SS;Price, MA;Crooks, GM
通讯作者：
Crooks, GM