Department of Infection, Immunity and Biochemistry, School of Medicine, Cardiff University, Cardiff, UK.
Haematologica. 2010 Mar;95(3):514-7. doi: 10.3324/haematol.2009.014381.
Studies of the role of individual genes in chronic lymphocytic leukemia (CLL) have been hampered by the inability to consistently transfect primary tumor cells. Here, we describe a highly efficient method of genetically modifying primary CLL cells using a VSVG pseudotyped lentiviral vector. We transduced CD38 negative CLL cells with a lentiviral vector encoding CD38 which caused increased surface CD38 expression in all the samples tested (n=17) with no evidence of plasmacytoid differentiation. The mean percentage of positive cells expressing CD38 was 87%+/-8.5% and the mean cell viability 74%+/-17%. This high level of transduction of all the CLL cell samples tested demonstrates the utility of this technique which should prove applicable for the introduction and analysis of other genes in these non-dividing cells.
个体基因在慢性淋巴细胞白血病(CLL)中的作用研究一直受到无法稳定转染原代肿瘤细胞的阻碍。在此,我们描述了一种使用 VSVG 假型慢病毒载体高效修饰原代 CLL 细胞的方法。我们用编码 CD38 的慢病毒载体转导 CD38 阴性的 CLL 细胞,在所有测试的样本(n=17)中导致表面 CD38 表达增加,没有浆细胞样分化的证据。表达 CD38 的阳性细胞的平均百分比为 87%+/-8.5%,细胞活力的平均百分比为 74%+/-17%。所有测试的 CLL 细胞样本的高转导效率证明了该技术的实用性,这应该适用于在这些非分裂细胞中引入和分析其他基因。