Kit Summary

For the expansion of human CD3+CD56+ Cytokine-induced Killer (CIK) 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+ CIK cells
  • Can use PBMCs as the starting cell population
  • Kit-expanded CIK cells are cytotoxic
  • Does not require specialized instrumentation
 

 

Why Expand CIK Cells Ex Vivo?

Cytokine-induced killer (CIK) cells exhibit phenotypic and functional similarities to both T cells and Natural Killer (NK) cells. While CIK cells express CD3 and expand readily in culture like T cells, they do not require functional priming for in vivo activity, a feature shared with NK cells. CIK cells consist of a heterogeneous population, including CD3+CD56-, CD3+CD56+, and CD3-CD56+ cells. Importantly, CD3+CD56+ have the most potent cytotoxic function among the CIK cells and express a number of cytokines, including IL-2, IFN-γ, TNF-α, and GM-CSF. The advantages that CIK cells have over other routes of cell therapy include their ease of in vitro propagation and their ability to be primed without the need for exogenous administration of IL-2. Over the past decade, cell therapy using CIK cells has emerged as an active area of research for the treatment of various types of cancer, such as hepatocellular carcinoma, non-small cell lung cancer, renal cell carcinoma, and gastric cancer. The ability to expand CIK cell populations ex vivo is valuable to researchers who require CIK 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 CIK cells.

  • Human Killer Cell Base Media 1
  • Recombinant Human IFN-γ
  • Recombinant Human IL-2
  • Mouse Anti-Human CD3
  • Reconstitution Buffer 1
  • Reconstitution Buffer 2

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 Cytokine-induced Killer Cells.
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Flow Cytometry Analysis of Expanded Human Cytokine-induced Killer Cells. Human PBMCs were cultured ex vivo for 21 days to expand Cytokine-induced Killer (CIK) cells using reagents included in the CellXVivo Human CIK Cell Expansion Kit. Compared to unexpanded PBMCs (A), PBMCs treated with the kit (B) show an increased number of CD3+CD56+ CIK cells. Flow cytometric analysis was performed on day 21 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 CIK Cells are Cytotoxic to Tumor Cells.
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Kit-expanded CIK Cells are Cytotoxic to Tumor Cells. Cytokine-induced Killer (CIK) cells expanded using the CellXVivo CIK Cell Expansion Kit were assessed for toxicity against the killer 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 CIK cells for 4 hours at the indicated effector-to-target cell (E:T) ratios (A). (B) Graph showing % killing of tumor cells by CIK cells at the indicated E:T ratios as determined by measuring relative fluorescence intensity of released Calcein-AM.

Preparation and Storage
  • Stability & 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.
Background: Natural Killer T (NKT) Cell Kits
  • Long Name:
    Natural Killer Cell Selection
  • Alternate Names:
    Natural Killer T (NKT) Cell Kits
Assay Procedure

Refer to the product datasheet for complete product details.

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

  • Isolate PBMC from human blood
  • Culture PBMCs in Human CIK Cell Expansion Media
  • Verify CIK cell expansion after 21 days in culture
 

 

Reagents Provided

Reagents Supplied in the CellXVivo Human CIK Cell Expansion Kit (Catalog # CDK014):

  • Human Killer Cell Base Media 1
  • Recombinant Human IFN-γ
  • Recombinant Human IL-2
  • Mouse Anti-Human CD3
  • Reconstitution Buffer 1
  • Reconstitution Buffer 2

 

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 CIK Cell Expansion using the CellXVivo Human CIK 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 CIK Cell Expansion Media. Culture the cells in the pre-coated 75 cm2 flask for a total of 14 days.

 

Culture the cells for a total of 21 days.

 

Refresh media and split cells on day 6, 12, and 18.

Culture the cells for a total of 21 days.

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

 

CIK cells are ready to be used for downstream applications.

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

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