There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.
Abstract
In this study, a fluorometric method using alamarBlue has been developed for detecting
cell-mediated cytotoxicity in vitro. AlamarBlue is a non-toxic metabolic indicator
of viable cells that becomes fluorescent upon mitochondrial reduction. Specific lysis
of targets by effector cells is quantified by comparing the total number of viable
cells in wells containing effector and targets together, with wells where target and
effector cells were separately seeded. Cell-mediated cytotoxic activity by alloreactive
T cells and natural killer cells has been detected using a novel application of the
alamarBlue technique. The assay that we have developed to detect cell-mediated cytotoxicity
is extremely sensitive and specific and requires a significant lower number of effector
cells than the standard 51Cr assay. Since alamarBlue reagent is non-toxic to cells
and the assay can be performed under sterile conditions, effector cells may be recovered
at the end for further analysis or cell expansion, if desired. Direct comparison of
cell-mediated cytotoxicity measured by the alamarBlue method with the standard 51Cr
release assay revealed that the former method is as specific and more sensitive than
the conventional assay. Moreover, very small inter and intra-assay variations have
been observed for alamarBlue cytotoxicity assays. In conclusion, this study shows
that the alamarBlue assay is an extremely sensitive, economical, simple and non-toxic
procedure to evaluate cell-mediated cytotoxicity that yields accurate results using
a limited number of effector cells. Furthermore, since this assay is a one-step procedure,
and does not involve any risk for the personnel, it may be useful to analyze automatically
cell-mediated cytotoxicity in a large number of samples.