T Cell FAQ
Q. Does PRIME-XV® T Cell CDM (chemical-defined media) contain any human-derived materials or materials that have been in contact with animal-derived materials during manufacturing?
A. No. T cell medium is a chemically-defined, animal component-free medium. Irvine Scientific CDM products do not contain any undefined animal-derived raw materials and are free of any animal-derived components, including human plasma or human blood-derived components. All raw materials are defined and their exact concentrations are known, which can include recombinant proteins.
Q. What is Irvine Scientific’s definition of animal component free (ACF)?
A. Irvine Scientific ACF products are manufactured with raw materials that do not contain any animal or human derived products to the tertiary level and are manufactured using equipment that does not come in contact with any animal or human products. The raw material handling, storage and final product manufacture and packaging are performed in dedicated human and animal component free facilities and equipment.
Q. What are the sources of T cells that were used to generate results in PRIME-XV® T Cell CDM?
A. Cryopreserved PBMCs from various donors were used to generate the expansion data on cell culture plate, bags, and G-Rex described in the brochure. Purified cryopreserved T cells were used for generating polarization/differentiation data. However, purified T cell expansion data is 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 there 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, our 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? Th or Treg?
A. This media will support the expansion of enriched T cell populations. Currently, we have data that indicates that Th1 and Treg 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. We also offer custom media development optimization for customers interested in enriching specific T cell populations.
Q. What density of cells do you recommend to maintain cells?
A. Here is our recommended protocol; however, our media can support higher cell densities (5 million cells/mL on G-Rex).
Q. How do we monitor changes in growth, since the media is devoid of Phenol red?
A. During development of the media, we tested cell viability and fold expansion of CD3+ (cell count) to monitor changes in growth. If Phenol Red is desired in the media, please contact us.
Q. Why do cells perform differently in PRIME-XV® T Cell CDM when compared to RPMI + 10% FBS?
A. By removing the serum and other undefined components the media is optimized specifically to deliver consistent vigorous growth for T cells and maintain T cell functionally and potency. Please see our blog post, The Impact of Serum and Serum- Derived Undefined Biologic Modifiers in Cell Culture for more information.
Q. Which reactors have been used? G-Rex, Wave etc.
A. Irvine Scientific has data available for expansion of T cells from PBMCs in G-Rex and culture bags. Please refer to the product insert for the G-Rex protocol and data.
Q. Does Irvine Scientific have any data to support transduction using PRIME-XV® T Cell CDM?
A. There are many factors that can affect transduction efficiency, which include donor variability, vector type, MOI (multiplicity of infection), and the customer’s specific culture and transduction protocols. When comparing PRIME-XV T Cell CDM with other benchmark media, we recommend the customers to evaluate the specific expansion profile obtained with our media for optimal results.
Q. How does the performance of PRIME-XV® T Cell CDM compare to other commercially available media?
A. Please see the data in this 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 the effecting the populations of expanded cells. While one lot can perform well for one cell type, it may be a poor performed 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, Cell, Media and Process - The Three Musketeers of Cell Culture for more information.
Q. How did we activate cells?
A. Coated cell culture plates or bags with anti-human CD3 and anti-human CD28 antibodies were used to activate the cells. Please refer to the product insert for the protocol. Soluble anti-CD3 and anti-CD28 antibodies were used for activation and expansion in the G Rex system.
Q. Does PRIME-XV® T Cell CDM contain antibiotics?
A. No. PRIME-XV T Cell CDM does not contain any antibiotics.
Q. What is Irvine Scientific’s policy for disclosing formulation information?
A. Different levels of details can be shared depending on the components and Irvine Scientific will work with our customers to address their formulation inquiries on a case by case base. Please contact us with your formulation requests.
Q. What is the maximum production batch size?