TY  - JOUR
TI  - A Feeder Cell-Free System for Chimeric Antigen Receptor Gene Transduction Into Natural Killer Cells
AU  - Kubo, Nobuhiro 
AU  - Baba, Minori 
AU  - Suzuki, Yuko 
AU  - Kasahara, Yasushi 
AU  - Hosokai, Ryosuke 
AU  - Imamura, Masaru 
AU  - Saitoh, Akihiko 
AU  - Imai, Chihaya 
VL  - 16
IS  - 8
PY  - 2026
DA  - 2026/04/20
SP  - e5669
C1  - Bio-protocol 2026;16:e5669
DO  - 10.21769/BioProtoc.5669
UR  - https://doi.org/10.21769/BioProtoc.5669
AB  - Anti-CD19 chimeric antigen receptor (CAR)-natural killer (NK) cells are expected to demonstrate anti-CD19 CAR-T-cell-like efficacy against relapsed and refractory B-cell malignancies and autoimmune diseases, with fewer adverse events and the added advantage of permitting the use of allogeneic cells. However, the methodology for generating CAR-NK cells remains under development. Although various cell sources and expansion methods are available, feeder cells derived from cancerous tissue have been most commonly employed to promote ex vivo expansion of NK cells. In the protocol described herein, NK cells are expanded from adult peripheral blood mononuclear cells using CD2- and NKp46-specific stimulating antibodies in combination with multiple cytokines. The activated NK cells can be genetically modified using a retroviral vector. Subsequent culture of these cells yields large numbers of anti-CD19 CAR-NK cells. The current method, which enables feeder-free, large-scale generation of anti-CD19 CAR-NK cells, eliminates the risk of tumor cell contamination and may facilitate safer clinical application.
KW  - NK cell expansion
KW  - CAR-NK cells
KW  - Anti-NKp46 antibody
KW  - Anti-CD2 antibody
KW  - Interleukin-12
KW  - Interleukin-18
KW  - Interleukin-21
KW  - Off-the-shelf
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False