A mouse fibroblast cell line, L929, was used to evaluate cell adhesion onto the t-hm-AlGltn films. The L929s were first cultured in a medium (RPMI-1640 (R8758, Sigma-Aldrich, St. Louis, MO, USA)) containing 2 v/v % fetal bovine serum. The t-hm-AlGltn films were placed on 24-well plates and a glass ring was put on each film. L929 cells (5.0 × 104 cells) were seeded onto each film for 5 min and the films were rinsed with 2 mL of phosphate buffered saline (PBS, pH 7.4). Then the cells were fixed with 10% formalin neutral buffer solution for 60 min and permeabilized in 0.2 v/v % Triton-X 100 in PBS for 2 min followed by 0.1% DAPI in PBS for 10 min in light-shielding conditions at room temperature. The adhered cells were observed with an IX81 inverted fluorescence microscope (Olympus Co. Ltd., Tokyo, Japan). The counted number of adhered cells was calculated from the area of the microscopic field (n = 3).
The cells were then observed with a scanning electron microscope (SEM). In brief, the cells were gradually dehydrated with a 50–99 v/v % ethanol/water solution. Then, the cells were immersed in tert-butylalcohol twice followed by freeze drying at −80 °C. The cells were then observed by SEM.
The cells were then observed with a scanning electron microscope (SEM). In brief, the cells were gradually dehydrated with a 50–99 v/v % ethanol/water solution. Then, the cells were immersed in tert-butylalcohol twice followed by freeze drying at −80 °C. The cells were then observed by SEM.