Journal: Life

Article Title: Effect of Butein, a Plant Polyphenol, on Apoptosis and Necroptosis of Prostate Cancer Cells in 2D and 3D Cultures

doi: 10.3390/life15060836

Figure Lengend Snippet: Cell viability, cell morphology, and nuclear morphology of butein against prostate cancer cell lines. ( A ) Chemical structure of butein. Formula: C 15 H 12 O 5 ; MW: 272.3. PC-3, DU145, and HPrEC cells were treated with butein at various concentrations (0, 1.25, 2.5, 5, 10, 15, 20, and 30 μM) and then cultured for 48 h. Viable cells were subjected to ( B ) trypan blue exclusion assay, and ( C ) MTT assay. ( D ) Cell morphology of PC-3 and DU145 cells was examined by phase contrast microscopy. ( E ) Nuclear morphology of PC-3 and DU145 cells was examined by fluorescence microscopy after DAPI (2 μg/mL) staining. All data are expressed as mean ± standard error of the mean of three independent experiments. *, p < 0.05 was compared to respective controls. HPrEC: human prostate epithelial cell line; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; and Conc., concentration.

Article Snippet: HPrEC cells were cultured in Prostate Epithelial Cell Base Medium (ATCC, PCS-440-030) supplemented with (ATCC, PCS-440-040).

Techniques: Cell Culture, Trypan Blue Exclusion Assay, MTT Assay, Microscopy, Fluorescence, Staining, Concentration Assay

Journal: Journal of Extracellular Biology

Article Title: Evaluating the Influence of Different Serum‐Free Culture Conditions on the Production and Function of Natural Killer Cell‐Derived Extracellular Vesicles

doi: 10.1002/jex2.70049

Figure Lengend Snippet: NK cell characteristics after different serum‐free culture conditions. (A) Workflow for static and shaking NK‐92 cultures, and purification of EVs. (B) Viability of NK‐92 cells after 48 h of culture in serum‐free media under static or shaking conditions supplemented with IL‐15 was assessed by propidium iodide by flow cytometry. Control samples were NK‐92 cells grown in Advanced RPMI medium with 20% FBS. Data are presented as the mean ± SD of four separate experiments. Statistical analysis performed using the Friedman one‐way ANOVA test indicated no statistical difference between the different culture conditions. (C) Expression levels of CD56 on NK‐92 cells after 48 h of culture in serum‐free media under static or shaking conditions. One representative experiment of three.

Article Snippet: NK‐92 cells (5 × 10 7 cells) were resuspended in 65 mL of serum‐free media with 10 ng/mL of recombinant human IL‐15 (R&D Systems) and cultured for 48 h. Six different serum‐free media optimized for culturing of NK cells were tested: CellGenix GMP SCGM (xeno‐free, Sartorius [abbreviated GMP SCGM]), CTS NK‐Xpander Medium (xeno‐free, ThermoFisher [abbreviated CTS]), Immunocult NK Cell Base Medium (xeno‐free, StemCell Technologies, abbreviated Immunocult), ExCellerate Human NK Cell Expansion Media, Xeno‐Free (R&D Systems, abbreviated ExCellerate XF), and ExCellerate Human NK Cell Expansion Media, Animal Component Free (R&D Systems, abbreviated ExCellerate ACF).

Techniques: Purification, Flow Cytometry, Control, Expressing

Journal: Journal of Extracellular Biology

Article Title: Evaluating the Influence of Different Serum‐Free Culture Conditions on the Production and Function of Natural Killer Cell‐Derived Extracellular Vesicles

doi: 10.1002/jex2.70049

Figure Lengend Snippet: TEM images of purified EVs derived from different serum‐free NK cell culture media. (A) TEM images of EVs obtained from static culture conditions. (B) TEM images of EVs obtained from shaking culture conditions. Scale bar 200 nm. Images are representative of 8–10 images from each sample.

Article Snippet: NK‐92 cells (5 × 10 7 cells) were resuspended in 65 mL of serum‐free media with 10 ng/mL of recombinant human IL‐15 (R&D Systems) and cultured for 48 h. Six different serum‐free media optimized for culturing of NK cells were tested: CellGenix GMP SCGM (xeno‐free, Sartorius [abbreviated GMP SCGM]), CTS NK‐Xpander Medium (xeno‐free, ThermoFisher [abbreviated CTS]), Immunocult NK Cell Base Medium (xeno‐free, StemCell Technologies, abbreviated Immunocult), ExCellerate Human NK Cell Expansion Media, Xeno‐Free (R&D Systems, abbreviated ExCellerate XF), and ExCellerate Human NK Cell Expansion Media, Animal Component Free (R&D Systems, abbreviated ExCellerate ACF).

Techniques: Purification, Derivative Assay, Cell Culture