Immuno-Oncology Advances in Applied Cellular Analysis - 25

Advances in Applied Cellular Analysis * Continuous Live-Cell Analysis of the Immune-Tumor Axis

ClearView plates and monitored for migration
via phase-contrast images (10x) collected automatically every 30 min for 24h using IncuCyte.
Addition of CXCL12 (3-200nM) in the basechamber caused clear concentration, and timedependent chemotaxis; higher concentrations
caused more rapid movement than lower concentrations and the measured EC50 was 23nM.
No directional migration was observed with
vehicle control or the inactive chemokine,
CXCL11 (100nM). Co-addition of the CXCR-4
receptor antagonist AMD3100 abolished the
response to CXCL12. Similar observations were
made with the human T-cell line, Jurkat. In four
assay plates run in parallel, the Z' values for the
high and low control groups ranged from 0.5-0.8
and CXCL12 EC 50 values from 19-33nM,
indicating high assay precision and
reproducibility. From the time-lapse videos,
the movement of individual T-cells through
the pores could be clearly observed.

Summary
For each of the immuno-oncology examples
provided, IncuCyte® CLCA yielded robust,
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Figure 7 (C & D).
Scale bar 100μM. (C) Note the time-and
SDF-1α concentration-dependent
chemotactic cell migration quantified as
a reduction in cell area on the topside of
the plate. (D) Concentration response
curves for SDF-1α obtained from
four separate experiments illustrates
assay reproducibility.

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Immuno-Oncology Advances in Applied Cellular Analysis

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