Natural Killer Cells: Methods and Protocols (Methods in Molecular Biology)

1. Tracking Effector and Memory NK Cells During MCMV Infection
Aimee M. Beaulieu and Joseph C. Sun

2. Application of Mass Cytometry (CyTOF) for Functional and Phenotypic Analysis of Natural Killer Cells
Alexander W. Kay, Dara M. Strauss-Albee, and Catherine A. Blish

3. Assessment of NK Cell Metabolism
Molly P. Keppel and Megan A. Cooper

4. Genotyping Single Nucleotide Polymorphisms and Copy Number Variability of the FCGRs Expressed on NK Cells
Amy K. Erbe, Wei Wang, Mikayla Gallenberger, Jacquelyn A. Hank, and Paul M. Sondel
5. Measurement of Average Telomere Length in Ex Vivo Expanded Natural Killer Cells by Fluorescence In Situ Hybridization (FISH)
Sourindra N. Maiti
6. In Vitro Assessment of Human Natural Killer Cell Migration and Invasion
Karin Tomin, Ronald H. Goldfarb, and Per Albertsson
7. Microfluidic-Based Live-Cell Analysis of NK-Cell Migration In Vitro
Saravanan Nandagopal, Francis Lin, and Sam K.P. Kung
8. Microwell-Based Live Cell Imaging of NK Cell Dynamics to Assess Heterogeneity in Motility and Cytotoxic Response
Bruno Vanherberghen, Thomas Frisk, Elin Forslund, Per E. Olofsson, Karolin Guldevall, and Björn Önfelt
9. Assessment of Natural Killer Cell Cytotoxicity Using Image Cytometry Method
Kelsey J. McCulley and Srinivas S. Somanchi
10. Analysis of Intracellular Ca²⁺ Mobilization in Human NK Cell Subsets by Flow Cytometry
Jakob Theorell and Yenan T. Bryceson
11. Using NK Cell Lipid Raft Fractionation to Understand the Role of Lipid Rafts in NK Cell Receptor SignalingEsther Serrano-Pertierra and Carlos López-Larrea
12. High- and Super-Resolution Microscopy Imaging of the NK Cell Immunological Synapse
Emily M. Mace and Jordan S. Orange
13. The Planar Lipid Bilayer System Serves as a Reductionist Approach for Studying NK Cell Immunological Synapses and their Functions
Grant Bertolet and Dongfang Liu
14. Expansion of NK Cells Using Genetically Engineered K562 Feeder Cells
Minh-Trang Thi Phan, Seung-hwan Lee, Sang-Ki Kim, and Duck Cho

15. Ex-Vivo Expansion of Human NK Cells Using K562 Engineered to Express Membrane Bound IL21
Srinivas S. Somanchi and Dean A. Lee

16. Large-Scale Culture and Genetic Modification of Human Natural Killer Cells for Cellular Therapy Natalia Lapteva, Robin Parihar, Lisa A. Rollins, Adrian P. Gee, and Cliona M. Rooney
17. Gene Modification of Human Natural Killer Cells Using a Retroviral VectorJoshua N. Kellner, Conrad R. Cruz, Catherine M. Bollard, and Eric S. Yvon
18. Modification of Expanded NK Cells with Chimeric Antigen Receptor mRNA for Adoptive Cellular TherapyYaya Chu, Allyson Flower, and Mitchell S. Cairo
19. mRNA Transfection to Improve NK Cell Homing to TumorsEmily Levy, Mattias Carlsten, and Richard Childs
20. In Vitro Generation of Human NK Cells Expressing Chimeric Antigen Receptor through Differentiation of Gene-Modified Hematopoietic Stem CellsEmily Lowe, L

This volume contains collection of Natural Killer Cell methodologies relevant for both basic and translational research. These methodologies present new developments in the natural killer (NK) cell field, such as understanding the influence of NK cells metabolism on its function, identifying complexity of NK cell subsets through mass cytometry, and determining the emergence of memory NK cells in murine model of MCMV infection. Methods that study NK cell migration and cytotoxicity through endpoint analysis or live single cell imaging are also discussed. Chapters also describe methods pertaining to translational application of NK cells, such as ex vivo expansion of NK cells on K562 cell lines genetically modified to express either membrane bound IL-15 or membrane bound IL-21, large scale NK cell culture, current techniques for engineering NK cells to express chimeric antigen receptors or chemokine receptors using retroviral vectors, electroporation of mRNA, and the natural phenomenon of trogocytosis. Written in the highly successful Methods in Molecular Biology series format, these chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Cutting edge and thorough, Natural Killer Cells: Methods and Protocols is a valuable resource for researchers who not only want to understand mechanisms that govern NK cell behavior and diversity, but also for those who want to understand how to systematically evaluate NK cells for adoptive immunotherapy applications.