CellXVivo Human NK Cell Expansion Kit

For the expansion of human CD3-CD56+ NK Cells from peripheral blood mononuclear cells (PBMCs)
Catalog # Availability Size / Price Qty
CDK015
Human NK Cell Expansion Using the CellXVivo™ Kit is Equivalent to Standard Feeder Cell Expansion. 
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Product Details
Procedure
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CellXVivo Human NK Cell Expansion Kit Summary

Kit Summary

For the expansion of human CD3-CD56+ NK Cells from peripheral blood mononuclear cells (PBMCs).

Key Benefits

  • Contains high quality bioactive cytokines and antibodies
  • Provides optimized reagents needed induce expansion of CD3-CD56+ NK cells
  • Can use PBMCs as the starting cell population
  • Kit-expanded NK cells are cytotoxic
  • Does not require specialized instrumentation
 

 

Why Expand NK Cells Ex Vivo?

Natural Killer (NK) cells were identified as a distinct sub-population of lymphocytes that have the capacity to spontaneously lyse tumor cells. Human NK cells are phenotypically characterized by the expression of CD56 and the absence of CD3. NK cells produce immunoregulatory cytokines, including interferon-γ (IFN-γ), tumor necrosis factor-beta (TNF-β), tumor necrosis factor-α (TNF-α), granulocyte macrophage-colony stimulating factor (GM-CSF), interleukin (IL)-10, and IL-13. Activation of NK cells can be triggered via NKp46 and NKp30 receptors or after stimulation with combinations of IL-2, IL-12, and IL-15. NK cells play an important role in both the adaptive and innate immune responses that govern infection, autoimmunity, and tumor immunosurveillance. Preclinical evidence and early clinical success has established NK cell immunotherapy as a promising therapeutic strategy in cancer. The ability to expand NK cell populations ex vivo is valuable to researchers who require NK cells for downstream applications including studies for cell therapy and cancer immunotherapy.

 

 

Reagents Provided

This kit contains the following reagents for the ex vivo expansion of human NK cells.

  • Human Killer Cell Base Media 2
  • NK Cell Expander 1
  • NK Cell Expander 2
  • NK Cell Expander 3
  • NK Cell Expander 4
  • NK Cell Expander 5
  • Reconstitution Buffer 1
  • Reconstitution Buffer 2
  • 20X Wash Buffer

Note: The components for this kit require different storage/shipping temperatures and will arrive in separate packaging.

Stability and Storage

Store the unopened kit at < -20 °C. Do not use past the kit expiration date.
*Provided this is within the expiration date of the kit.

Precaution

When handling biohazardous materials such as human cells, safe laboratory procedures should be followed and protective clothing should be worn.

Limitations

  • FOR LABORATORY RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.
  • The safety and efficacy of this product in diagnostic or other clinical uses has not been established.
  • This reagent should not be used beyond the expiration date indicated on the label.
  • Results may vary due to variations among cells derived from different donors.
 

 

Data Examples
Flow Cytometry Analysis of Expanded Human NK Cells.
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Flow Cytometry Analysis of Expanded Human NK Cells. Human PBMCs were cultured in vitro for 14 days to expand Natural Killer cells using reag ents included in the CellXVivo Human NK Cell Expansion Kit. Compared to unexpanded PBMCs (A), PBMCs treated with the kit (B) show an increased number of CD3-CD56+ NK cells. Flow cytometric analysis was performed on day 14 of the expansion and cells were stained with Human NCAM-1/CD56 PE-conjugated Antibody, and Human CD3ε PerCP-conjugated Antibody. Quadrants were set based on isotope-stained samples.

Kit-expanded NK Cells are Cytotoxic to Tumor Cells.
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Kit-expanded NK Cells are Cytotoxic to Tumor Cells. NK Cells expanded using the CellXVivo NK Cell Expansion Kit were assessed for toxicity against the NK cell-sensitive K562 tumor cell line. K562 cell were loaded with the live-cell dye, Calcein-AM (Tocris, Catalog # 5119) prior to being mixed with NK cells for 4 hours at the indicated effector-to-target cell (E:T) ratios (A).

Human NK Cell Expansion Using the CellXVivo Kit is Equivalent to Standard Feeder Cell Expansion.
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Human NK Cell Expansion Using the CellXVivo Kit is Equivalent to Standard Feeder Cell Expansion. Human NK cells were enriched from peripheral blood mononuclear cells and expanded (3 x 106 starting population) using the CellXVivo NK Cell Expansion Kit or the standard method using artificial antigen-presenting K562 clone 9.mbIL-21 cells. NK cell expansion was quantified at days 1, 7, 10, and 14. The rate of NK cell expansion observed using the CellXVivo Kit was similar to that observed using K562 cells. Data courtesy of the laboratory of Dr. Bruce Walcheck at the University of Minnesota.

Human NK Cells Expanded using the CellXVivo Kit Express High Levels of CD16.
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Human NK Cells Expanded using the CellXVivo Kit Express High Levels of CD16. Human NK cells were enriched from peripheral blood mononuclear cells and expanded ( 3 x 106 starting population) using the CellXVivo NK Cell Expansion Kit or by the standard method using artificial antigen-presenting K562 clone 9.mbIL-21 cells. After 14 days of expansion, NK cells were assessed for CD16 expression. NK cell expanded using the CellXVivo Kit expressed higher levels of CD16 based on mean fluorescence intensity when compared to NK cells expanded using the K562 method. Data courtesy of the laboratory of Dr. Bruce Walcheck at the University of Minnesota.

Human NK Cells Expanded using the CellXVivo Kit Express NK Cell-specific Markers.
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Human NK Cells Expanded using the CellXVivo Kit Express NK Cell-specific Markers. Human NK cells were enriched from peripheral blood mononuclear cells and expanded (3 x 106 starting population) using the CellXVivo NK Cell Expansion Kit (A, C, E) or by the standard method using artificial antigen-presenting K562 clone 9.mbIL-21 cells (B, D, F). After 14 days of expansion, NK cells were assessed for expression of characteristic NK cell surface markers, CD56 (A, B), NKG2D (C, D), and NKp46 (E, F). Data courtesy of the laboratory of Dr. Bruce Walcheck at the University of Minnesota.

Specifications

Shipping Conditions
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.
Species
Human

Product Datasheets

Data Example

Human NK Cell Expansion Using the CellXVivo™ Kit is Equivalent to Standard Feeder Cell Expansion.  Human NK cells were enriched from peripheral blood mononuclear cells and expanded (3 x 106starting population) using the CellXVivoNK Cell Expansion Kit or the standard method using artificial antigen-presenting K562 clone 9.mbIL-21 cells. NK cell expansion was quantified at days 1, 7, 10, and 14. The rate of NK cell expansion observed using the CellXVivoKit was similar to that observed using K562 cells.Data courtesy of the laboratory of Dr. Bruce Walcheck at the University of Minnesota.

Kit-expanded NK Cells are Cytotoxic to Tumor Cells. NK Cells expanded using the CellXVivoNK Cell Expansion Kit were assessed for toxicity against the NK cell-sensitive K562 tumor cell line. K562 cell were loaded with the live-cell dye, Calcein-AM (Tocris, Catalog # 5119) prior to being mixed with NK cells for 4 hours at the indicated effector-to-target cell (E:T) ratios (A).

Flow Cytometry Analysis of Expanded Human NK Cells.  Human PBMCs were culturedin vitrofor 14 days to expand Natural Killer cells using reag ents included in the CellXVivoHuman NK Cell Expansion Kit. Compared to unexpanded PBMCs (A), PBMCs treated with the kit (B) show an increased number of CD3-CD56+NK cells. Flow cytometric analysis was performed on day 14 of the expansion and cells were stained with Human NCAM-1/CD56 PE-conjugated Antibody, and Human CD3ε PerCP-conjugated Antibody. Quadrants were set based on isotope-stained samples.

Human NK Cells Expanded using the CellXVivo™ Kit Express NK Cell-specific Markers.  Human NK cells were enriched from peripheral blood mononuclear cells and expanded (3 x 106starting population) using the CellXVivoNK Cell Expansion Kit (A, C, E) or by the standard method using artificial antigen-presenting K562 clone 9.mbIL-21 cells (B, D, F). After 14 days of expansion, NK cells were assessed for expression of characteristic NK cell surface markers, CD56 (A, B), NKG2D (C, D), and NKp46 (E, F). Data courtesy of the laboratory of Dr. Bruce Walcheck at the University of Minnesota.

Human NK Cells Expanded using the CellXVivo™ Kit Express High Levels of CD16. Human NK cells were enriched from peripheral blood mononuclear cells and expanded ( 3 x 106starting population) using the CellXVivoNK Cell Expansion Kit or by the standard method using artificial antigen-presenting K562 clone 9.mbIL-21 cells. After 14 days of expansion, NK cells were assessed for CD16 expression. NK cell expanded using the CellXVivoKit expressed higher levels of CD16 based on mean fluorescence intensity when compared to NK cells expanded using the K562 method.Data courtesy of the laboratory of Dr. Bruce Walcheck at the University of Minnesota.

CellXVivo Human NK Cell Differentiation Kit

Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, CD3-CD56+ Natural Killer (NK) cells are expanded ex vivo from peripheral blood mononuclear cells with the CellXVivo Human NK Cell Expansion Kit using the following procedure:

  • Isolate PBMC from human blood
  • Culture PBMCs in Human NK Cell Expansion Media
  • Verify NK cell expansion after 14 days in culture
 

 

Reagents Provided

Reagents Supplied in the CellXVivo Human NK Cell Expansion Kit (Catalog # CDK015):

  • Human Killer Cell Base Media 2
  • NK Cell Expander 1
  • NK Cell Expander 2
  • NK Cell Expander 3
  • NK Cell Expander 4
  • NK Cell Expander 5
  • Reconstitution Buffer 1
  • Reconstitution Buffer 2
  • 20X Wash Buffer

 

Other Supplies Required

Reagents

  • Ficoll-Hypaque

Equipment

  • 75-cm2 cell culture flasks
  • Microscope
  • Hemocytometer
  • 37 °C and 5% CO2 humidified cell culture incubator
  • Centrifuge
  • Pipettes and pipette tips
  • Sterile deionized water

 

Procedure Overview

Protocol for NK Cell Expansion using the CellXVivo Human NK Cell Expansion Kit.

Isolate PBMCs from human blood.

Isolate PBMCs from human blood.

Perform a cell count.

Perform a cell count.

Suspend 1 x 106 PBMCs/mL in Human NK Cell Expansion Media.

 

Culture the cells for a total of 14 days.

 

Refresh media and split cells on day 4, 7, and 10.

Culture the cells for a total of 21 days.

Verify expanded NK cells by analyzing NK cell marker expression via flow cytometry (optional).

 

NK cells are ready to be used for downstream applications.

Verify expanded NK cells by analyzing NK cell marker expression via flow cytometry (optional)

Reagents Provided

FAQs

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