Key Performance Indicators
Successful proliferation of NK cells from various sources has been achieved both with and without feeder cells, without IL-2, and at low (2-3% v/v) serum levels
*1,2
NK cells cultured in Enable-NK™ Duo media exhibited a biomarker-determined cellular phenotype much closer to freshly-isolated NK cells
*1,2,3
Levels of biomarkers for cellular exhaustion (i.e. PD-1, TIM-3, LAG-3, TIGIT, NKG2A) were reduced by about 20-25%, upon NK cells being cultured in Enable-NK™ Duo cell culture media
*2,3
NK cells cryopreserved in Enable-NK™ Duo demonstrated better post-thaw viability, and improved CD16 retention with recovery of ADCC function (ADCC: Antibody Dependent Cell-mediated Cytotoxicity)
*1,2
2-3 times improved retroviral transduction efficiency has been reported with Enable-NK™ Duo cell culture media
*1
Enable-NK™ Duo can support reduction in serum dependence, down to 2-3% v/v serum content
*1,3
As described in the table on the Products page, Enable-NK™ Duo requires minimal cytokine supplementation
*1
NK Cells cultured in Enable-NK™ Duo demonstrated elevated NAD values compared to cells cultured in control media
*1
NK Cells cultured in Enable-NK™ Duo participated in more NK cell signaling and recruitment of other immune cells (e.g. T cells) through the secretion of cytokines such as Interferon-gamma (IFN-γ)
*1
NK cells cultured in Enable-NK™ Duo media demonstrated enhanced antifungal potency against Cryptococcus neoformans
*4
Enable-NK™ Duo cell culture media is not reliant on extensive cytokine supplementation
*1,2
30-40% (or higher) improvement in cytotoxic performance against cancer cells, by NK cells expanded in Enable-NK™ Duo cell culture media
*1,2,3
Comparison Chart
Enable-NK™ Duo | Competitor 1 | Competitor 2 | Competitor 3 | |
---|---|---|---|---|
Serum concentration as low as 2%? | ||||
IL-2 concentration as low as 100 U/mL? | ||||
IL-12 is NOT required? | ||||
IL-15 is NOT required? | ||||
IL-21 is NOT required? | ||||
Can be added as a supplement to other media? |
Technical Documents
Good Manufacturing Practices
Good Manufacturing Practices (GMP) are an extensive set of regulations and guidelines that ensure a high standard of quality and consistency in the processes of manufacturing of ancillary materials and therapeutic products. These standards were established in 1969 and published as Part 128 of the Code of Federal Regulations (CFR).
Onkologic Inc. emphasizes the importance of quality throughout the process of manufacturing media products, and follows GMP guidelines for ingredients, materials, training, and manufacturing.
RESEARCH USE GRADE Vs GMP GRADE
Onkologic Inc. provides both Research-grade and GMP-grade cell culture media products, both grades manufactured using the same raw materials and identical processes, but with greater quality oversight and documentation for the GMP-grade products.
Research Use Grade vs GMP Grade
Quality Attributes | Research Use Only (RUO) | GMP |
---|---|---|
Manufacturing and release of products through QMS | Yes | Yes |
Batch records with full quality oversight | Adequate | Extensive |
Rigorous raw material and supplier qualification (USP <1043>) | Adequate | Extensive |
USP validated test methods | Yes | Yes |
Expiry date on Certificate of Analysis (CoA) | Yes | Yes |
Regulatory support | Limited | Yes |
Frequently Asked Questions (FAQs)
Yes, Enable-NK™ Duo can be ordered as either cell culture media (250 ml or 500 ml volumes each for Enable-NK™ Grow and Enable-NK™ Activate), or as Enable-NK™ Duo 10x - a 10x supplement that can be added to any preferred starting media of the user’s choice (25 ml or 50 ml volumes each for Enable-NK™ Grow 10x and Enable-NK™ Activate 10x). We recommend using the supplement at a 1:10 dilution into basal media.
Enable-NK™ Grow and Enable-NK™ Activate can be purchased as cell culture media in volumes of 250 ml or 500 ml each. Enable-NK™ Grow 10x and Enable-NK™ Activate 10x supplements can be purchased separately in volumes of 25 ml or 50 ml each.
Enable-NK™ Duo has been deliberately designed in two phases in order to achieve the two objectives of expansion and activation separately. Previously published protocols for NK cell expansion use a single-phase media formulation with cytokine additives to accomplish both proliferation and activation simultaneously, resulting in hyperactivation and exhaustion of NK cells. In contrast, we created two cell culture media formulations: Enable-NK™ Grow is optimized for proliferation and Enable-NK™ Activate is optimized for activation, with the purpose of accomplishing both objectives without hyperactivating or exhausting the expanded NK cells.
It is our recommendation that Enable-NK™ Grow be used prior to Enable-NK™ Activate, and that the transition from Grow to Activate is accomplished through dilution rather than centrifugation. At the same time, the user is free to customize either Enable-NK™ Grow or Enable-NK™ Activate in a manner that best fits their protocol and experimental goals. As an example, Grow and/or Activate can be used in isolation, but we do recommend using them sequentially (i.e. Grow followed by Activate). Please see downloadable guidelines and protocols for more details.
Yes. However, please keep in mind that Enable-NK™ Grow has been optimized for proliferation and optional serum reduction, while it does accomplish activation to some extent. Enable-NK™ Activate has been optimized for activation of NK cells, while it does support proliferation to some extent. We strongly recommend that Enable-NK™ Grow and Enable-NK™ Activate be used sequentially, per downloadable guidelines, instructions and protocols.
Our standard offering is Grow and Activate media in combination, but we can create custom orders to offer each phase individually. For custom orders, please reach out to us at contact@onkologic.com, or call us at 508-216-0009.
Enable-NK™ can accomplish serum-reduction through the use of its Enable-NK™ Grow formulation and taking a stepwise approach for serum content reduction over time. Please see downloadable guidelines, instructions and protocols for details.
We have consistently seen proliferation of NK cells being supported by Enable-NK™ media at as low as 2% v/v serum content.
Yes, Enable-NK™ media when used in combination with feeder cells, can provide synergy corresponding to proliferation and cytotoxic performance of expanded NK cells. Please refer to research posters available for download under Technical Documents on the Resources page of our website.
No. While Enable-NK™ can support feeder-free expansion of NK cells, the ideal use of Enable-NK™ is in combination with feeder cells in order to provide synergy for both proliferation and cytotoxicity of expanded cells.
Enable-NK™ media are fully customizable for any cytokine cocktail of the user’s choice. For internal development efforts, we only used interleukin-2 (IL-2) at a concentration of 100 U/mL. However, other levels of IL-2 (e.g. lower at 50 U/ml or higher at 200 U/ml) and combinations with other cytokines (e.g. IL-15 at 10 ng/ml) were effectively employed by collaborating NK cell research groups.
Enable-NK™ Grow is formulated with DMEM/F12 media as a starting point, and with the addition of a 20+ molecular ingredients (no plant extracts). Enable-NK™ Activate has the same formulation as Enable-NK™ Grow plus a few additional ingredients (including 7 plant extracts), in order to enhance the activation of NK cells
Yes, although Enable-NK™ Grow is optimized for proliferation.
Yes, although Enable-NK™ Activate is optimized for activation.
Yes. Supportive data is available in the downloadable research posters under Technical Documents on the Resources page of our website.
Yes. Supportive data is available in the downloadable research posters under Technical Documents on the Resources page of our website.
The following are some of the key attributes of NK cells which are enhanced with the use of Enable-NK™ cell culture media:
Key Performance Metrics
More technical information is available in the research posters that can be downloaded under Technical Documents on the Resources page of our website.
Based on a number of collaborative studies, 3 major streams of the underlying molecular mechanisms have emerged: