Large-Scale Culture and Genetic Modification of Human Natural Killer Cells for Cellular Therapy

Methods Mol Biol. 2016;1441:195-202. doi: 10.1007/978-1-4939-3684-7_16.


Recent advances in methods for the ex vivo expansion of human natural killer (NK) cells have facilitated the use of these powerful immune cells in clinical protocols. Further, the ability to genetically modify primary human NK cells following rapid expansion allows targeting and enhancement of their immune function. We have successfully adapted an expansion method for primary NK cells from peripheral blood mononuclear cells or from apheresis products in gas permeable rapid expansion devices (G-Rexes). Here, we describe an optimized protocol for rapid and robust NK cell expansion as well as a method for highly efficient retroviral transduction of these ex vivo expanded cells. These methodologies are good manufacturing practice (GMP) compliant and could be used for clinical-grade product manufacturing.

Keywords: G-Rex; Genetic modification with retroviral vector; Large-scale expansion; Natural killer cells.

MeSH terms

  • Blood Component Removal
  • Cell Culture Techniques / methods*
  • Cell Proliferation
  • Cell Transplantation
  • Feeder Cells / cytology
  • Feeder Cells / immunology
  • Feeder Cells / metabolism
  • Humans
  • K562 Cells
  • Killer Cells, Natural / cytology*
  • Killer Cells, Natural / immunology
  • Killer Cells, Natural / metabolism
  • Lymphocyte Activation
  • Transduction, Genetic*