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Natural Killer (NK) Cells for Cell Therapy

Natural killer cells have the potential to address common limitations of T cell-based immune cell therapies. NK cell biology offers promising advantages for both therapeutic activity and manufacturing simplification, leading to improved delivery of effective therapy to patients.

  • Reduced risk of cell therapy failure due to antigen escape. NK cells act by several mechanisms besides antigen specific CAR targeting. Their redirected cell lysis and antibody-directed cellular cytotoxicity (ADCC) functions serve as antigen-independent means to kill mutant target tumor cells.
  • Improved scalability and speed of therapy delivery. NK cells are amenable to developing allogeneic therapies based on universal cell banks. These banks can supply therapeutic cells much more rapidly than autologous processes which require patient cells to produce each treatment.
NK cell genetically engineered for immune cell therapy

As Your Program Advances Towards the Clinic, We Will Grow With You

Adherent cells in culture supported by extracellular matrix

Serum-Free and Animal-Free Cell Culture

Increase the consistency of your cell cultures as you approach translational studies for cell therapy programs. Adopting these media will:

  • Reduce variability in media composition
  • Simplify compliance with regulatory guidelines
  • Simplify comparability testing for raw material changes
gowned cleanroom operator carrying cell culture vessel

Transitioning From Research to GMP

As you advance from discovery into process development and clinical manufacturing, choose GMP raw materials with the equivalent performance as the research-grade products you use. This will make your transition as efficient and seamless as possible.

NK Cell Isolation

Begin your immunotherapy workflow on a strong footing. Accurate and reproducible isolation of NK cell populations will lay the groundwork for a successful cell therapy program. After cell isolation, confirm that your procedure is effective with thorough cell phenotype analysis.

cell surface receptors bound by antibodies for NK cell isolation
antibodies used for NK cell selection

NK Cell Selection

Antibody specificity is the basis for accurate and reproducible cell selection. Choosing the right antibodies will enable you to reliably isolate high purity NK cell populations.

View Custom Antibody Services

  • Recombinant antibody engineering and GMP antibodies
  • Monoclonal panel development
  • Specialty conjugation
induced pluripotent cells in culture for differentiation into NK cells

NK Cells From Induced Pluripotent Stem Cells

Differentiation of iPSCs into NK cells facilitates the generation of universal cell banks to treat multiple patients with reduced cost and more rapid development.

NK Cell Culture and Expansion

Generating enough NK cells to support clinical trials requires a consistent and scalable expansion protocol. To boost culture consistency, QC your media preparations with precise quantitation of cytokines and growth factors. Analyze your expanded cultures for cell number, viability, and functionality.

NK cell targeted by NK Cloudz for cell culture expansion
Antibodies

Cell Selection

Pro dots

Cytokines & Growth Factors

GMP ProDots Proteins are formulated as dissolvable spheres enclosed inside sterile, weldable bags for closed processing cytokine addition.

Animal-Free GMP Proteins are produced in dedicated controlled-access laboratories to prevent potential contamination by animal components or byproducts.

GMP small molecules

Small Molecules

GMP small molecules promote somatic cell reprogramming to iPSC prior to NK cell differentiation - our ancillary grade small molecules facilitate simple translation from research-use to GMP.

ScaleReady Logo

Bio-Techne believes in the power of partnerships. We have joined with Fresenius Kabi and Wilson Wolf to bring you ScaleReady™, lowering the barriers to entry into the immune cell therapy space for all sizes and stages of manufacturing programs. ScaleReady brings cell and gene therapies to life by offering a significant reduction of complexity and cost while consistently providing superior repeatability and cell quality.

The ScaleReady product portfolio supports scalable feeder-free NK cell expansion, delivering true platform, process, and product continuity for your NK cell programs. When paired with the Lovo® and Cue® functionally-closed-system automated cell processing systems, Bio-Techne reagents and the G-Rex® Bioreactor enable high throughput parallel processing of cell therapies within a small footprint.

NK Cell Engineering

Enhance the performance of your NK cell-based therapy and realize the potential of gene engineering. Partner with our experts for customized number of cells/vial, validation testing, datasheet preparation, and batch record documentation. We will work with you to make sure we engineer the right solution for your goals.

Gene engineering
TcBuster(TM) - A next-generation non-viral gene delivery system
Watch the video to see how TcBuster delivers your genes of interest.

TcBuster for Non-Viral Gene Delivery

Accomplish your NK cell engineering more easily by using non-viral gene delivery. TcBuster will revolutionize your ability to engineer the optimum phenotype for your cell therapy.

  • Reduced cost of genetic modification
  • Rapid cell engineering with manufacturing controls
  • Large cargo size integration
  • Flexibility to deliver multiple genes in one operation
  • Not subject to supply shortage of lentivirus vectors

NK Cell Engineering With Lentivirus

Vector characterization

Process efficiency improvements with viral transduction enhancers

 

 

Learn About Vector Characterization at ProteinSimple

AAV and lentiviral viral vector manufacturing solutions
analysis of NK cell phenotype after cell engineering

Confirming Successful Cell Engineering

Once you have engineered your NK cells, verify that their phenotype reflects the gene editing you intended.

flow cytometry analysis of NK cells after transduction with human CD19 CAR construct

Human NK cells were enriched from PBMCs and either (A) transduced or (B) not transduced with a human CD19 CAR. Cells were stained with Human CD56 PE-conjugated Antibody and Recombinant Human CD19 Protein, Atto 647N Conjugate.

NK Cell Characterization

Final product characterization of your NK cells should cover all critical quality attributes (CQAs) to ensure your product is as safe as possible for in vivo administration. Using orthogonal assays to measure unrelated parameters will enable thorough analysis of the cell surface, secretory profile, NK cell cytolytic activity, and residual contaminants.

characterization of NK cells for immune cell therapy
phenotype of NK cell for immune cell therapy

NK Cell Phenotype

Perform a detailed phenotypic characterization of your NK cells to determine if they express the right surface receptors for the cell subset and activation status you need. In addition, measure population heterogeneity and the absence of residual feeder cells from culture expansion.

analysis of NK cells with the FlowX Human NK Cell Phenotyping Kit

NK cell phenotypic analysis with the FlowX Human NK Cell Phenotyping Kit. NK cells were expanded from PBMCs in ExCellerate Human NK Cell Expansion Media with plate-bound anti-human NKp46 supplemented with recombinant human IL-2, IL-12, IL-18, and IL-21 for 13 days. Cells were analyzed with antibody conjugates included in the kit: NKp30  Alexa Fluor 488, CD16 PerCP, CD56 Alexa Fluor™ 647, NKG2D Alexa  Fluor 700, NKp46 PE, and CD3. Gating was based off live cells (using the Live-or-Dye 405/545 dead cell exclusion dye), and quadrants were set using isotype controls (not shown).

immunoassay detection of effector molecules released by activated NK cells

NK Cell Function

To make sure your cells are functional as well as phenotypically intact, assay their release of effector NK cell molecules including IFN-gamma, Granzymes, and Perforin. In addition, directly measure the ability of your cells to lyse target cells in co-culture.

analysis of NK cell cytotoxicity with the FlowX Human NK Cell Killing Flow Cytometry Kit

CAR-NK cell cytotoxicity using the FlowX Human NK Cell Killing Flow Cytometry Kit. Human NK cells were enriched from PBMCs and transduced with a human CD19 CAR (CAR-NK). CAR-NKs were labeled with Janelia Fluor® 646, Daudi target cells or K562 target cells were labeled with MitoMark Green, and cells were co-cultured for 2 hours at the indicated Effector to Target ratios. Anti-LAMP-1/CD107a PE antibody was included in the culture during the 2 hour killing assay to measure degranulation. Cells were harvested and stained with anti-human CD56 APC and CD3 Alexa Fluor 405 antibodies. To measure target cell death, CD3-/CD56- Janelia Fluor 646- cells were analyzed for MitoMark Green.

Micro-Flow Imaging: I See Particles
Watch the video to see how MFI distinguishes different types of particles in a cell culture sample.

Contaminant Screening

Make sure that your NK cell product is clean enough for in vivo administration, either in animal models or in clinical trials. Confirm that the levels of particulates, residual cell activation beads, and other impurities are low enough to meet your requirements.

ACD RNAscope in situ Hybridization (ISH) Technology Overview
Watch the video for an overview of RNAscope in situ hybridization technology.

Tissue Biopsy and NK Cell Localization

Monitor the biodistribution of your NK cell therapy in vivo with single-cell resolution in the context of tissue morphology.

  • RNAscope and BaseScope assays for quantitative transgene mRNA analysis in intact tissues
  • Good Clinical Laboratory Practice (GCLP)-compliant RNAscope ISH analysis for clinical and preclinical studies
  • Professional Assay Services for tissue-based expression analysis support
RNAscope Multiplex Fluorescent V2 Assay detection of NK cells in situ in cervical cancer

Detection of NK cells in human cervical cancer using the RNAscope Multiplex Fluorescent V2 Assay. NK cells were detected in situ with specific marker probes for NCR1, CD56, XCL1, and CCL5. Read the scientific poster for more details.

Custom Services

No cutting edge cell therapy should be delayed from reaching patients just because the right materials aren’t available. If you need specially designed reagents or assays for developing your NK cell therapy, we will work with you to deliver optimized solutions for streamlining your process. We will gladly provide customized formulation and packaging as well as bulk orders.