Q. Does PRIME-XV T Cell CDM contain any human-derived materials or materials that have been in contact with animal-derived materials during manufacturing?

A. PRIME-XV T Cell CDM is a chemically defined, animal component-free (ACF) medium. FUJIFILM Irvine Scientific CDM products do not contain any undefined raw materials. All raw materials are defined and their exact concentrations are known, including recombinant proteins. In addition, CDM products are manufactured using only human and ACF raw materials and ancillary materials in a dedicated ACF area.

Q. Does PRIME-XV T Cell CDM contain antibiotics?

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

Q. How does the performance of PRIME-XV T Cell CDM compare to that of other commercially-available media?

A. For examples of PRIME-XV T Cell CDM compared to other commercially-available media, please see the data in the product insert. PRIME-XV T Cell CDM was designed to perform without the addition of serum. The addition of human AB serum can affect the T cell phenotype due to lot-to-lot variability of human AB serum, which could potentially inhibit the desired phenotype. Different serum types and lots will impact monocytes, thus affecting the populations of expanded cells. While one lot can perform well for one cell type, it may be poor performance for others. A CDM avoids such variations between lots and will perform consistently for the desired cell type.

Optimal cell culture performance requires the cells, the media, and the process all work together closely in harmony. Irrespective of the end-use application, changes to any one of these aspects can significantly impact the culture and ultimately result in compromised performance. Optimization of the protocol is highly encouraged to achieve optimal culture performance with our media. Please refer to our blog post for more information, Cell, Media and Process - The Three Musketeers of Cell Culture.

Q. Do cells perform differently in PRIME-XV T Cell CDM when compared to RPMI + 10% FBS?

A. Serum-containing media can introduce variability to cell culture conditions and cell performance, as the contents of animal components can widely vary. As a chemically defined product, the contents and exact concentrations of PRIME-XV T Cell CDM's formulations are known. As a result, using PRIME-XV T Cell CDM ensures consistent, vigorous growth for T cells, maintains T cell functionality, and retains potency just as well or better than the serum-containing alternatives. More information can be found on the FUJIFILM Irvine Scientific blog post, The Impact of Serum and Serum-Derived Undefined Biologic Modifiers in Cell Culture.

Q. How are changes in growth monitored since the media is devoid of phenol red?

A. During development of the media, we tested cell viability and fold expansion of CD3⁺ cells (cell count) to monitor changes in growth. If phenol red is desired in the media, please contact FUJIFILM Irvine Scientific for custom media services.

Q. Does PRIME-XV T Cell CDM support transduction?

A. Yes, PRIME-XV T Cell CDM is proven to support transduction without the use of enhancers or spinoculation. Customers should evaluate the specific expansion profile obtained with the media for optimal results.

Q. What are the sources of T cells that were used to generate results for PRIME-XV T Cell CDM?

A. Both fresh and cryopreserved PBMCs derived from various donors were used to generate the expansion data on G-Rex and Quantum bioreactor described in the brochure. Expanded and re-stimulated T cells were used for generating polarization/differentiation data. Enriched CD3⁺ T cell expansion data is also available upon request.

Q. Are the original T cell fractions driven towards a specific T cell phenotype upon expansion? Are the CD4⁺ and CD8⁺ fractions maintained in similar ratios after expansion?

A. The ratio of CD4⁺ to CD8⁺ cells can be affected by a number of factors, including the donor from which they were derived, the presence of monocyte populations, the method of activation, etc. As a result, the CD4/CD8 ratio may be affected during or after expansion. While the ratio may change during culture, PRIME-XV T Cell CDM can support the expansion of both populations. Multiple donors were used in the development of the media.

Q. Has PRIME-XV T Cell CDM been tested on enriched cell populations? Th₁, Th₂, CD8⁺?

A. This media will support the expansion of enriched T cell populations. There is data demonstrating that TH₁, TH₂, and CD8⁺ cells are supported by this media when supplemented with appropriate cytokines. PRIME-XV T Cell CDM is especially suited for further optimization for specific T cell populations as its components are defined in comparison to serum-containing media. Custom media development and media optimization services are available for customers interested in enriching specific T cell populations.

Q. What is the maximum production batch size?

A. For more information on batch size and supply chain capabilities, please contact FUJIFILM Irvine Scientific.

Q. Is the use/addition of any supplements recommended to enhance growth in PRIME-XV T Cell CDM or PRIME-XV T Cell Expansion XSFM?

A. FUJIFILM Irvine Scientific does not currently offer any supplements for PRIME-XV T Cell CDM or PRIME-XV T Cell Expansion XSFM. CDM from FUJIFILM Irvine Scientific is typically not supplemented.

Q. Are any cytokine cocktails used for the culture? If so, in what concentration?

A. The protocol has been developed and validated with our media containing 300 IU/mL of IL-2. The recommended concentrations of IL-7, IL-15, and IL-21 are being evaluated.

Q. Can T cells be grown with PRIME-XV T Cell CDM in the complete absence of IL-2?

A. There is some data on T cell growth in the absence of interleukins in PRIME-XV T Cell CDM. On average, CD3⁺ fold expansion is 20-fold to 40-fold less, and viability is 70-80% at day 10 compared to > 90% in IL-2-containing conditions.

Q. How were the cells activated?

A. Soluble anti-CD3 and anti-CD28 antibodies were used for activation and expansion in the G-Rex and Quantum systems. Please refer to the product insert for the protocol.

Q. Is an adaptation step needed for shifting to a PRIME-XV T Cell Expansion XSFM or PRIME-XV T Cell CDM if the cells were previously cultured/stored in serum-containing media or a different media?

A. An adaptation step is not needed for shifting to a PRIME-XV T Cell Expansion XSFM or PRIME-XV T Cell CDM.

Q. What is the density of cells that can be supported by PRIME-XV T Cell CDM?

A. See our recommended protocol; however, our media can support higher cell densities up to 5.0x10⁶ cells/mL on G-Rex and 3.0x10⁶ cells/mL on Quantum.

Q. Which bioreactors have been used to culture the T cells using PRIME-XV T Cell CDM?

A. FUJIFILM Irvine Scientific has data available for expansion of T cells from PBMCs in G-Rex and Quantum. Please refer to the product insert for the G-Rex and Quantum protocol and data.

Q. What is the preferred culture platform to use PRIME-XV T Cell CDM? (12-well or 6-well plates, dishes, flasks, G-Rex, Quantum, Prodigy)

A. G-Rex, Quantum, and culture bags are the preferred culture platforms for PRIME-XV T Cell CDM. Please refer to the product insert for the G-Rex and Quantum protocol and data. For use in plates and flasks, reach out to Technical Support Services for more information.

Q. At what temperature is the T cell media in bags kept when operating a bioreactor?

A. After removal from refrigerated conditions, the bags may be kept hanging near the bioreactor at room temperature until completion of cell culture application. The current protocol reports successful performance and function without the need for refrigeration of media bags throughout operation.