eoe sub articles

EoE Sub Articles

All procedures involving human participants have been performed in compliance with the institutional, national, and international guidelines for human welfare and have been reviewed by the local institutional review board.
1. Preparation of materials
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	<li>Store reagents, antibodies, and chemicals as shown in the Material Safety Data Sheet (MSDS). Dissolve the drugs or cytokines in solvents as stock solutions and then aliquot for storage at -20 &#176;C or -80 &#176;C.<br...

an in vitro assay to study the cytotoxic capability of cytokine-induced killer cells

Source: Hsiao, C. H., et al. <a href="https://app.jove.com/v/60420">Isolation and Expansion of Cytotoxic Cytokine-induced Killer T Cells for Cancer Treatment.</a> J. Vis. Exp. (2020).
This video demonstrates an in vitro technique to measure the cytotoxic ability of cytokine-induced killer (CIK) cells against cancer cells. Upon co-culturing target cancer cells with CIK cells, the dead cancer cells are identified using a fluorescent viability dye and are quantified using flow cytometry, estimating the cytotoxic effect of CIK cells.

<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/21802/21802_Figure1.jpg" /> 
Figure 1: The proportion of CD3+CD56+ T cells from a representative PBMC sample. (A) Lymphocytes were recognized by specific size and granularity. Selected single cell population for analysis by flow cytometry. (B) Statistical analysis of CIK expansion efficacy from three healthy donors was conducted using a t-test (*, p &#60; 0.01).
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An In Vitro Assay to Study the Cytotoxic Capability of Cytokine-Induced Killer Cells

All procedures involving human participants have been performed in compliance with the institutional, national, and international guidelines for human ...

Take a culture of cancer cells with fluorescently labeled cytoplasmic proteins.
Add cytokine-induced killer or CIK cells &#8212; effector T cells with natural killer cell-like characteristics &#8212; possessing anti-tumor activity.
Natural killer group 2 member D or NKG2D&#160; receptors on the CIK cells bind to stress-inducible proteins on the cancer cells.
The binding induces CIK cells to release cytoplasmic granule toxins &#8212; perforins and granzymes.
Perforins form pores in the cancer cell membrane, allowing the entry of granzymes, which activate caspases, inducing apoptosis.
Post-incubation, harvest the cells. Add a fluorescent nucleic acid-binding cell viability dye.
The dye enters through the damaged cell membrane and binds to DNA, resulting in dual-stained dead cells.
With an intact cell membrane, live cells are impermeable to the dye.
Pass the cells through a flow cytometer to detect live cells displaying single fluorescence and dead cells with dual fluorescence.
Create a scatter plot to measure the percentage of dead cancer cells, quantifying the cytotoxic effect of CIK cells.