Journal: Biochemistry and Biophysics Reports

Article Title: Low level electricity increases the secretion of extracellular vesicles from cultured cells

doi: 10.1016/j.bbrep.2019.100713

Figure Lengend Snippet: Particle sizes and ζ-potentials of the EVs harvested from each cell line.

Article Snippet: A murine melanoma cell line, B16F1, was purchased from DS Pharma Biomedical Co., Ltd (Osaka, Japan).

Techniques:

Journal: Biochemistry and Biophysics Reports

Article Title: Low level electricity increases the secretion of extracellular vesicles from cultured cells

doi: 10.1016/j.bbrep.2019.100713

Figure Lengend Snippet: Particle distribution and morphology of the EVs collected from culture cells. B16F1 and 3T3 Swiss Albino cells were treated by low level electricity (ET; 0.34 mA/cm 2 ) for 1 h. After 24 h of incubation, EVs were collected from the conditioned medium of each cell line by ultracentrifugation. Then, the particle distributions of the EVs collected from B16F1 ((A): ET (−), (B): ET (+)) and 3T3 Swiss Albino ((C): ET (−), (D) ET (+)) were analyzed with a qNano. The morphologies of the EVs collected from B16F1 ((E): ET (−), (F): ET (+)) and 3T3 Swiss Albino ((G): ET (−), (H): ET (+)) were observed by TEM. Scale bars = 100 nm.

Article Snippet: A murine melanoma cell line, B16F1, was purchased from DS Pharma Biomedical Co., Ltd (Osaka, Japan).

Techniques: Incubation

Journal: Biochemistry and Biophysics Reports

Article Title: Low level electricity increases the secretion of extracellular vesicles from cultured cells

doi: 10.1016/j.bbrep.2019.100713

Figure Lengend Snippet: Enhancement of EV secretion from cultivated cells by ET. The particle number of the collected EVs from B16F1 (A) and 3T3 Swiss Albino (B) cells treated with or without low level electricity (0.34 mA/cm 2 , 1 h) was determined. The data show the mean ± S.D. (n = 5). * P < 0.05 vs. ET (−). The representative graphs of the particle concentration curve were obtained from B16F1- (C) and Swiss 3T3 Albino-derived EVs (D).

Article Snippet: A murine melanoma cell line, B16F1, was purchased from DS Pharma Biomedical Co., Ltd (Osaka, Japan).

Techniques: Concentration Assay, Derivative Assay

Journal: Biochemistry and Biophysics Reports

Article Title: Low level electricity increases the secretion of extracellular vesicles from cultured cells

doi: 10.1016/j.bbrep.2019.100713

Figure Lengend Snippet: ET increased the amount of protein in the collected EVs. The quantity of protein in the EVs harvested from the conditioned medium of B16F1 (A) and 3T3 Swiss Albino (B) was measured by BCA assay. The number of cells was counted by trypan blue staining after collecting their conditioned media. The relative percentage of viability to that of non-treated cells is indicated (C: B16F1, D: 3T3 Swiss Albino). The data show the mean ± S.D. (n = 5). * P < 0.05, ** P < 0.01 vs. ET (−).

Article Snippet: A murine melanoma cell line, B16F1, was purchased from DS Pharma Biomedical Co., Ltd (Osaka, Japan).

Techniques: BIA-KA, Staining

Journal: Biochemistry and Biophysics Reports

Article Title: Low level electricity increases the secretion of extracellular vesicles from cultured cells

doi: 10.1016/j.bbrep.2019.100713

Figure Lengend Snippet: Western blot analysis of the expression of the exosomal marker proteins. EVs derived from B16F1 (A) and 3T3 Swiss Albino (B) were subjected to SDS-PAGE (1 μg/lane). Western blotting was performed to observe the expression of exosome markers CD9, HSP70, and CD81. Per the product datasheet, the detected molecular weights of CD9, HSP70, and CD81 are approximately 25, 70–78, and 22–26 kDa, respectively. The experiments were performed 3 times independently. The ratios of particle number per protein amount of the EVs collected from B16F1 (C) and 3T3 Swiss Albino (D) were calculated based on the results of , .

Article Snippet: A murine melanoma cell line, B16F1, was purchased from DS Pharma Biomedical Co., Ltd (Osaka, Japan).

Techniques: Western Blot, Expressing, Marker, Derivative Assay, SDS Page

Journal: Biochemistry and Biophysics Reports

Article Title: Low level electricity increases the secretion of extracellular vesicles from cultured cells

doi: 10.1016/j.bbrep.2019.100713

Figure Lengend Snippet: Effect of Rho GTPase inhibition on increase in EV secretion by ET. B16F1 and 3T3 Swiss Albino cells were exposed to ET (0.34 mA/cm 2 ) for 1 h in the presence of Rho GTPase inhibitor at the indicated concentration. After 24 h of additional incubation, EVs were collected, and the particle number of the collected EVs from B16F1 (A) and 3T3 Swiss Albino (B) cells was then measured with a qNano. The relative particle number of each group to that in control (ET (−)) are presented. The data show the mean ± S.D. (n = 4). Significant difference; * P < 0.05.

Article Snippet: A murine melanoma cell line, B16F1, was purchased from DS Pharma Biomedical Co., Ltd (Osaka, Japan).

Techniques: Inhibition, Concentration Assay, Incubation, Control

Journal: Biochemistry and Biophysics Reports

Article Title: Low level electricity increases the secretion of extracellular vesicles from cultured cells

doi: 10.1016/j.bbrep.2019.100713

Figure Lengend Snippet: Cellular uptake of EVs collected with or without exposure of the cells to ET. B16F1 and 3T3 Swiss Albino cells (1 × 10 5 cells/35-mm dish) were incubated with PKH67-labeled EVs (5 μg protein/mL) derived from B16F1 and 3T3 Swiss Albino cells, respectively, for 6 h at 37 °C. Then, the cells were fixed, and their nuclei were counterstained with DAPI. Fluorescence images of B16F1 (A and B) and 3T3 Swiss Albino (D and E) were obtained by confocal laser scanning microscopy. Blue and green colors show the fluorescence of DAPI (nuclei) and PKH67 (EVs), respectively. The images of right columns are merged images of those of DAPI, PKH67, and phase contrast. Scale bars = 50 μm. The relative fluorescence intensities of EVs (ET (+)) to that of EV (ET (−)) was calculated from 12 images per each group of B16F1 (C) and 3T3 Swiss Albino cells (F) for one experiment by using Image J. The experiments were performed three times independently. The data show the mean ± S.D. (n = 3).

Article Snippet: A murine melanoma cell line, B16F1, was purchased from DS Pharma Biomedical Co., Ltd (Osaka, Japan).

Techniques: Incubation, Labeling, Derivative Assay, Fluorescence, Confocal Laser Scanning Microscopy