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Natural Killer (NK) and CAR-NK Cell Expansion Method using Membrane Bound-IL-21-Modified B Cell Line
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1Department of Pathology, Immunology and Laboratory Medicine,Rutgers-New Jersey Medical School, 2Department of Microbiology, Biochemistry & Molecular Genetics, Public Health Research Institute Center, New Jersey Medical School,Rutgers University, 3Center for Immunity and Inflammation, New Jersey Medical School,Rutgers-The State University of New Jersey
Chapters
- 00:04introduction
- 00:54Natural Killer (NK) Cell Expansion from Liver Tissues: Day 0
- 03:14Primary Natural Killer (NK) Cell Expansion from Peripheral Blood Mononuclear Cells (PBMCs): Day 0
- 04:34Attachment of 293T Cells (Day 2) and Retrovirus Transfection (Day 3)
- 05:58Retronectin Plate-Coating (Day 3) and Transduction (Day 4)
- 08:18CAR-NK Cells Collection (Day 6 or 7)
- 09:04Results: Expansion of PBNK Cells and Flow Cytometric Analysis of CAR-NK Cells
- 10:22Conclusion
Here, we present a method to expand peripheral blood natural killer (PBNK), NK cells from liver tissues, and chimeric antigen receptor (CAR)-NK cells derived from peripheral blood mononuclear cells (PBMCs) or cord blood (CB). This protocol demonstrates the expansion of NK and CAR-NK cells using 221-mIL-21 feeder cells in addition to the optimized purity of expanded NK cells.
