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March 09, 2017

In vitro T-cell proliferation assay

Background:

A useful assay to analyze the immunomodulatory potential of a test item is to analyze T-cell proliferation in vitro using the Jurkat cell line. This in vitro method is an easy and reproducible tool which does not require PBMC isolation and T-cell preparation. Jurkat cells, a human mature leukemic cell line, phenotypically resemble resting human T lymphocytes and has been widely used to study T cell physiology.

Method:

In vitro tests for T-cell proliferation assess the response of cells in culture to the test item at different concentrations. The presence of any cytotoxic effect is indicated by loss of cell viability, while the increased proliferation suggests a positive immunomodulatory effect of the test material.

In the in vitro proliferation assay based on the Jurkat cell line, the extent of cell proliferation is evaluated with a quantitative method based on photometric measurements. The Tetrazolium salt WST-1, is metabolically reduced in viable cells to water-soluble formazan. Thus the number of viable cells correlates to the colour intensity determined by photometric measurements.

Outcome:

As the number of viable cell correlates to the colour intensity determined by absorbance values at 450 nm, a proliferation % can be calculated to characterize the possible effect of a test item on T-cell proliferation.

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