Figure 5

Release of A549 cells from the hydrogels. Hydrogels were polymerized and freeze-dried as described earlier and decorated with YFP-LecB. Cells were seeded into the hydrogels and incubated for 2 h to guarantee cell adhesion. (A) Cell loaded hydrogels were incubated with different concentrations of L-fucose and about 90% of cells could be released with 100 µM of L-fucose. (B) YFP-LecB after elution of the cells. Flow cytometry analysis was conducted, revealing that about 50% of all cells still bear YFP-LecB on the surface. (C) Cell-loaded gels were incubated with 100 µM of L-fucose, samples were taken at regular intervals and cells were counted. After approx. 10 min, 90% of the cells were eluted from the matrix. (D) Biocompatibility of materials and procedures used. Cells were eluted from the hydrogel and cytotoxicity was investigated with flow cytometry. (E) Cells were seeded and grown for 3 days in the hydrogels, eluted with 100 µM of L-fucose and counted with a Neubauer counting chamber to see proliferation of cells. (F) Cells were eluted from the hydrogel with 100 µM of L-fucose and re-cultured under 2D conditions over 4 days (left side) and the growth was compared with a fresh control (right side). (G) Adhesion of eluted cells after being re-cultured in 2D. The right side shows the average surface area of re-cultured cells compared to a fresh control. The right side shows the actin filament of re-cultured and fresh cells to observe possible differences in their morphology. All bars represent the standard deviation. The significance was tested with a one-way ANOVA with alpha = 0.05 for Fig. 5A, E and G and with a two-way ANOVA for Fig. 5F with alpha = 0.05.