Q. Is PRIME-XV NK Cell CDM a chemically defined and animal component-free medium?

A. PRIME-XV NK Cell CDM is a chemically defined and animal component-free medium. A chemically defined medium is where all chemical components have known composition and concentration (e.g. recombinant proteins). This formulation provides reproducible results, traceability, and regulatory compliance.

Q. Does PRIME-XV NK Cell CDM contain any cytokines or growth factors?

A. PRIME-XV NK Cell CDM is a complete basal media and does not contain cytokines or growth factors. Supplementation of cytokines will depend on the particular application and is up to the end-user to determine.

Q. Does PRIME-XV NK Cell CDM contain any glutamine?

A. PRIME-XV NK Cell CDM contains glutamine.

Q. Does PRIME-XV NK Cell CDM contain antibiotics, phenol red, or some other pH indicator?

A. PRIME-XV NK Cell CDM does not contain antibiotics, phenol red, or any other pH indicator. However, if there is a need to incorporate antibiotics or phenol red in the media, custom media services are available. Please contact FUJIFILM Irvine Scientific for more information.

Q. What types of feeder cells were used for the expansion of NK cells with PRIME-XV NK Cell CDM? Were the feeder cells genetically modified?

A. Feeder cells were not used for the expansion of NK cells using PRIME-XV NK Cell CDM.

Q. What cell sources were used to expand NK cells with PRIME-XV NK Cell CDM?

A. Commercially-available freshly isolated peripheral blood mononuclear cells (PBMCs) were used.

Q. When PBMCs were used as a starting material, were NK and NKT cells enriched prior to expansion?

A. Positive selection for CD56⁺ or a negative selection for non-NK cell depletion was applied. It depended on the experimental goal; however, the cells expanded comparably with either isolation method.

Q. Were any cytokines used for the expansion of NK cells with PRIME-XV NK Cell CDM?

A. The following cytokines were used to expand NK cells with PRIME-XV NK Cell CDM: IL-2 (500 IU/mL), IL-12 (10 ng/mL), IL-18 (10 ng/mL), and IL-21 (10 ng/mL). To expand NKT cells, IL-2 was added at 1000 IU/mL.

Q. Has PRIME-XV NK Cell CDM been tested with any immortalized cell lines?

A. Protocol and data are available upon request through Technical Support Services.

Q. Is adaptation required when transitioning from a serum-containing media to PRIME-XV NK Cell CDM?

A. For PBMC-derived NK and NKT cell expansion, cells were expanded in PRIME-XV NK Cell CDM directly. No adaptation to serum-free conditions was necessary. However, if an increase in doubling time or impact in other cell culture parameters, a sequential adaptation into lower serum-containing conditions should be considered. This is particularly relevant if cells have previously been cultured in media with serum for multiple passages before being transitioned to PRIME-XV NK Cell CDM.

Q. What is the maximum cell density that PRIME-XV NK Cell CDM can support?

A. Up to 5.0x10⁶ cells/mL have been cultured successfully in PRIME-XV NK Cell CDM. See the recommended protocols in the product insert. However, the absolute maximum cell density has not been determined.

Q. What vessels were used for the expansion of NK cells with PRIME-XV NK Cell CDM?

A. NK cells have been expanded with PRIME-XV NK Cell CDM in tissue culture plates, flasks, and the G-Rex bioreactor.