Rt ces

A real-time cell electronic sensing system (RT-CES) (Wako Pure Chemical, Osaka, Japan) was used to analyze the cell adhesion assay as described^{48 (link)}. Microelectronic cell sensor arrays were integrated into the bottom of the microplates (E-Plate (16X) code 583-84881, model number DC16-B-NT) (ACEA Biosciences, Inc., San Diego, CA, USA). The sensor provides information that increases electrical resistance (cell index) to indicate an increase in cell adhesion. E-Plates were coated with collagen-1 (CL-1) (5 μg/m in PBS, 100 μL/well) at room temperature (RT) for 1 h and blocked with 1% BSA/10% FBS in D-MEM (100 μL/well) at RT for 1 h. After blocking the wells, S1 or V4 cells (1 × 10⁴) were seeded in each well of 16-well plates containing culture medium. Then, exosomes (0.5, 2.0 and 4.0 μg) derived from V4 cells were added to S1 cells, and exosomes (0.5, 2.0 and 4.0 μg) derived from S1 cells were added to V4 cells at 0 h or 1 h. Control cells were not treated with exosomes. Simultaneously, a cell adhesion assay was performed by adding exosomes from S1 cells and anti-GD2 mAb or adding exosomes from S1 cells, anti-GD2 mAb and anti-integrin β1 mAb to V4 cells at 0 h. Changes in cell adhesion were monitored continuously and are expressed as the cell index (CI).

Real-time cell electronic sensing system (RT-CES) (Wako Pure Chemical, Osaka, Japan), was used for analysis of cell adhesion and expansion. Changes in the biological status of cells can be measured automatically in real time by RT-CES. Microelectronic cell sensor arrays are integrated into the bottom of the micro-plates (E-Plate (16X) (ACEA Biosciences Inc., San Diego, CA)). This sensor provides continuous and quantitative information concerning the biological status of the cells in the well and the increased electrical resistance (Cell Index) indicates the increase of cell adhesion and expansion. E-Plate was coated by CL-1, CL-IV or FN at 5 μg/mL in PBS (100 μL/well) at room temperature for 1 h, and blocked by 1% BSA/D-MEM (100 μL/well) at room temperature for 1 h. After blocking of the plate, cells (1 x 10⁴) were added in each well of the plate containing culture medium and changes in cell adhesion and expansion were monitored continuously.