Haematologica (Mar 2010)

Genetic modification of primary chronic lymphocytic leukemia cells with a lentivirus expressing CD38

  • Laurence Pearce,
  • Liam Morgan,
  • Thet Thet Lin,
  • Saman Hewamana,
  • R. James Matthews,
  • Silvia Deaglio,
  • Clare Rowntree,
  • Christopher Fegan,
  • Christopher Pepper,
  • Paul Brennan

DOI
https://doi.org/10.3324/haematol.2009.014381
Journal volume & issue
Vol. 95, no. 3

Abstract

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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.