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Application: Primary NK cell engineering using viral transduction.
Background: NK (natural killer) cells are part of the innate immune system. They function in a histocompatibility complex-independent mode and derive from the hematopoietic lineage. They are the first line of defense against cancer. Expression of marker CD56 correlates with NK cell functionality: the CD56bright subset accounts for about 5% of the population and are less cytotoxic than the CD56dim subset. Cytotoxicity can happen by the release of perforin and granzyme, while activation by KARs (killer activating receptors) leads to release of Fas Ligand, TRAIL (TNF-related apoptosis-inducing ligand) and TNFα (tumor necrosis factor-alpha). In a suppressive tumor microenvironment, NK cells can become inhibited and unable to fight cancer cells. Several clinical trials have focused on using ex vivo generated NK cells alone or in combination with other approaches. NK cells can be generated ex vivo from peripheral blood, umbilical cord blood, iPS cells or immortalized NK cell lines. The ability to generate a number of pure cells high enough for human dosage often requires the use of growth factors such as IL-2 (interleukin 2) or IL-15, and feeder cells.CAR (chimeric antigen receptor) engineering can add target specific cytotoxicity to NK cells. CAR-NK cells typically have enhanced cytotoxic capacity and cytokine production through co-stimulatory molecules. They may provide stronger tumor-specific targeting and cytotoxicity than CAR-T cells. The use of CAR (chimeric antigen receptor)-NK cells is an expanding area holding great promise in cancer therapy.
Description: The Primary NK Transduction Kit is a complete kit suitable for primary NK cell engineering using viral transduction. It contains engineered Growth-Arrested NK Feeder Cells (#78912), Frozen Human Peripheral Blood NK Cells (#78798), NK Medium, Serum-Free (#82615), NK Cell Culture Cytokine Cocktail (#82616), and NK Viral Transduction Enhancer (#82617). The kit was optimized using high titer anti-CD19 CAR lentiviruses (such as BPS Bioscience #78601). The kit provides enough components for transduction of a starting population of more than 1 million NK cells.
Storage Stability: NK Cells and Feeder Cells are shipped in dry ice and should immediately be thawed or stored in liquid nitrogen upon receipt. Do not use a -80°C freezer for long term storage. NK Medium, Serum-Free, NK Cell Culture Cytokine Cocktail, and Viral Transduction Enhancer (Components A and B) are shipped in dry ice and stored at -20°C. After the first thaw, the medium can be stored at 4°C for up to 1 month. Alternatively, aliquot the medium into tubes and store at -20°C. Do not re-freeze aliquots after thawing.
Warnings: Avoid freeze/thaw cycles
Biosafety Level: Not applicable (BSL-1)
References: Du N., et al., 2021 Cancers (Basel) 13 (16): 4129.
Liu E., et al., 2020 N Engl J Med. 382(6):545-553.
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