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?

A.

Irvine Scientific has the capability of manufacturing up to 5,000 L batch size for PRIME-XV® T Cell CDM. Irvine Scientific can also do custom formulations. Please contact us for more information.

Q.

Is PRIME-XV® MSC Expansion SFM serum-free, animal-component-free?

A.

PRIME-XV MSC Expansion SFM is serum-free. However, it is not animal-component-free.

Q.

Can I use the PRIME-XV® MSC Expansion SFM to culture cells without a coating substrate?

A.

No, it is not recommended. Unlike serum-containing MSC culture medium, serum-free culture condition for human MSCs requires fibronectin (such as PRIME-XV Human Fibronectin, Catalog # 31002) or our proprietary matrix protein (PRIME-XV MatrIS F, Catalog # 31001). Without a coating substrate, MSCs will die in absence of serum.

Q.

What are the advantages of PRIME-XV® MSC Expansion SFM?

A.

PRIME-XV MSC Expansion SFM outperforms leading competitors in cell expansion studies while maintaining MSC gross morphology, cell surface expression markers, multipotency potential and immune modulation abilities. Additionally, PRIME-XV MSC Expansion SFM has been tested to support optimal cell expansion of different sources of MSCs, including bone marrow-derived and adipose-derived MSCs. Little to no adaptation is required when shifting from a serum-containing medium to our medium. PRIME-XV MSC Expansion SFM is ready-to-use with no additional growth supplements required and is supplied in a convenient one 250mL complete component.

Q.

Why switch over from existing products or serum-containing medium?

A.

Our medium has been shown to outperform 10% serum-containing medium and leading competitors in human MSC cell expansion studies. Cell growth is more optimal in our medium, without compromising its immune modulation functions or phenotypic characteristics. Therefore, end-users will spend fewer resources (culture medium, labor, plastic-ware, and ancillary reagents) to culture MSCs. PRIME-XV® MSC Expansion SFM does not contain serum. This eliminates the lot-to-lot variability often associated with serum.

Q.

What are the main components in PRIME-XV® Hypothermic Preservation Solution and PRIME-XV Cryogenic Preservation Solution?

A.

Both PRIME-XV Hypothermic Preservation Solution and PRIME-XV Cryogenic Preservation Solution formulations are proprietary. However, they are serum-free, protein-free aqueous solutions containing various sugars, salts and buffers to provide optimal pH buffering. PRIME-XV Cryogenic Preservation Solution also contains 10% DMSO. All raw material components are USP/ multi-compendial or highest available grade, including water for injection (WFI) quality water and were made under GMP condition.

Q.

Is it normal to see precipitation in the PRIME-XV® MSC Expansion SFM?

A.

As human source material is present in the product it may develop some precipitate matter during storage. It is suggested to remove the precipitates from the media through filtration or centrifugation at 1500 rpm for 5 minutes after thawing the media. This filtration step does not affect product performance.

Q.

Why do we see a 400-fold expansion for adipose-derived MSCs while there is only an 18-fold increased for bone marrow-derived MSCs when using PRIME-XV® MSC Expansion SFM?

A.

MSCs derived from different sources, i.e. the origin of where the cells were isolated from, have different intrinsic growth rate. Regardless of the MSC sources, both populations of cells showed higher expansion rates in PRIME-XV MSC EXPANSION SFM when compared to 10% serum-containing medium.

Q.

What does xeno-free mean?

A.

A xeno-free product contains no animal-derived component but may potentially contain human-derived components.

Q.

What does animal component-free mean?

A.

An animal component-free product contains neither animal nor human-derived materials.

Q.

How often are MSCs passaged when cultured in PRIME-XV® MSC Expansion SFM?

A.

Cells are often passaged when they reach 80% confluency, or the coverage of the culture vessel by the cells. One hundred percent confluency refers to cells covering 100% of the cultured surface area. Time between each passage depends on the media performance and initial seeding density. However, on average, human mesenchymal stem cells (MSCs) cultured in PRIME-XV MSC EXPANSION SFM is passaged every 3-4 days.

Q.

Can PRIME-XV® MSC Expansion SFM formula be customized according to the customer’s needs?

A.

Yes. We can modify our formula and create a custom batch. For more information, please email us at tmrequest@irvinesci.com.

Q.

Can I use the PRIME-XV® cell therapy products for clinical applications?

A.

Our PRIME-XV product line is intended for research use or for further manufacturing use only. PRIME-XV products are not for use in diagnostic procedures. The safety and efficacy of these products in diagnostic or other clinical uses has not been established. However, we are able to generate custom, clinical-grade culture media using our existing scalable industrial manufacturing capabilities. Please contact us directly at tmrequest@irvinesci.com for further inquiries.

Q.

Where can I find more information on which PRIME-XV® products have DMFs (Drug Master File)?

A.

Please contact us at tmrequest@irvinesci.com and reference the product name and catalog number for which you are inquiring about DMFs.