In vitro response of Cellero's negatively selected purified human T Cells in the presence of allogeneic antigen-presenting cells (APC).
Cryopreserved T cells offered by Cellero are purified from peripheral blood mononuclear cells by negative selection. The purity of our T cell products are confirmed for each lot by flow cytometric analysis. Furthermore, these T cells can be used in an in vitro assay directly after thawing.
Our featured T cells (lot no. 2817AP15) were tested for the magnitude of in vitro response to allogeneic stimuli by incubating them in the presence of several types of antigen-presenting cells (APC) from an allogeneic donor.
The extent of T cell proliferation were measured using the flow cytometry-based method of labelling responder cells (T cells) with 5,6-carboxyfluorescein diacetate succinimidyl ester (CFDA-SE), which is converted to fluorescent 5,6-carboxyfluorescein succinimidyl ester (CFSE) inside the cells and covalently labels intracellular proteins. When labelled cells divide, half of the labelled proteins are distributed to each daughter cell, resulting in a decrease in dye intensity and a peak that is shifted to the left on a histogram of CFSE fluorescence intensity.
Figure 2 below incorporates proliferation data from each sample with different APC type and number. It allows for a comparison of the capacity the different allogeneic APC to stimulate CD4+ T cell response. The data clearly shows that mature DC are superior to immature DC and monocytes in stimulating an in vitro allogeneic response.