2.2. High aspect ratio vessel
Rotatory cell culture system (RCCS-1) was purchased from Synthecon, Incorporated (Houston, TX 77054, USA) with Autoclavable High Aspect Ratio Vessel (HARV) of 50 ml capacity. Spores from 7 days old culture were used. Approximately 1012 Laquinimod were added to the HARV and rotated at 25 rpm in the horizontal axis, inside the chamber with 90% humidity and maintained at 25 °C.
Polyurethane, 10 wt% (PU, Estane® Skythane® X595A-11, Lubrizol) was prepared by dissolving overnight in a solvent mixture of N,N dimethylformamide (DMF, Samchun, Korea) and methyl ethyl ketone/2-butanone (MEK, extra pure, Samchun, Korea) (50/50, wt:wt%). 2 wt% the dextran (from Leconostoc mesenteroides, average MW = 8500–11,500, Sigma–Aldrich) has been added to the solution along with culture filtrate. The obtained solutions were placed in a plastic syringe tube and fed through a metal capillary (nozzle) with a diameter di = 0.21 mm (21 G) attached to a 1-D robot-system basal body moves laterally and is controlled by the LabVIEW 9.0 software program (National Instrument). The feeding rate was maintained at 0.5 ml/h via a controllable syringe pump. Electrospinning was carried out at a voltage of 18 kV and working distance of 15 cm at room temperature.