Characteristics of fluorescent phagokinetic assay. (A) Transwell cell motility assay. A greater number of U118MG cells (1.33 fold) transit through the filter compared with T98G. Graph depicts the mean number of cells that transit through the filter of 5 10X fields in 4 replicate wells/cell line (20 fields/cell line/experiment) from 3 independent experiments; p = 0.0185). (B) Mean area of fluorescent particles cleared per cell reveals U118MG cell line exhibits greater intrinsic motility than T98G (2.29 fold greater area cleared/cell). Graph depicts the mean motility of cells from 3 independent experiments; 100 cells/cell line measured for each experiment; p = 0.0012. (C) Fluorescent phagokinetic assay reveals differences in motility of cell lines (primary mouse cerebral cortical astrocytes, T98G, U118MG) on different extracellular matrices (fibronectin (FN), type IV collagen, laminin). * indicates p < 0.001 comparing motility of U118MG on FN vs. Collagen IV or FN vs. laminin. For all pairwise comparisons of U118MG on any substratum vs. either T98G or primary astrocytes, p < 0.001. (D) Histogram of distribution of areas cleared by U118MG vs. T98G from a representative experiment on poly-D-lysine treated tissue culture plastic (no additional substratum). Results are expressed as means +/- SEM, and statistical significance was evaluated by Student's t-test (A, B)or one-way ANOVA followed by Newman-Keuls post-hoc test (C).
Windler-Hart et al. BMC Cell Biology 2005 6:14 doi:10.1186/1471-2121-6-14