Journal: Cell Reports Methods

Article Title: 3D model for human glia conversion into subtype-specific neurons, including dopamine neurons

doi: 10.1016/j.crmeth.2024.100845

Figure Lengend Snippet:

Article Snippet: For 10x Genomics snRNA-seq, single nuclei suspensions were loaded onto 10x Genomics Single Cell 3′ Chips along with the mastermix as per the manufacturer’s protocol ( https://support.10xgenomics.com/single-cell-gene-expression/index/doc/technical-note-chromium-single-cell-3-v3-reagent-workflow-and-software-updates ) for the Chromium Single Cell 3′ Library to generate single nuclei gel beads in emulsion (GEMs, version 3 chemistry).

Techniques: Recombinant, Protease Inhibitor, Fluorsave, SYBR Green Assay, Expressing, Software

Journal: Scientific Reports

Article Title: Dissecting multi drug resistance in head and neck cancer cells using multicellular tumor spheroids

doi: 10.1038/s41598-019-56273-6

Figure Lengend Snippet: Comparative drug response profiles of 2D cell cultures and MCSs. ( a – c ) Drug response curves for LK0917, LK0902, and LK1108 for 2D and MCSs cultures treated with 170–17000 µM of doxorubicin; 3.33–333 µM of cisplatin; and 2.2–220 µM of methotrexate. ( d ) IC 50 (µM) values calculated from the drug response curves for both 2D and MCSs of all cell types for all drugs. Significantly large differences between the IC 50 values of 2D and MCSs are denoted with “ # ”. Each IC 50 value is the average of three independent experiments (n = 3) with triplicates set for each concentration.

Article Snippet: For the generation of MCSs sized 300–500 μm, 200 μL of LK0917 (MCS 17 ), LK0902 (MCS 02 ), and LK1108 (MCS 08 ) single cell suspensions were seeded in ultra-low attachment (ULA) plates (Corning Life Sciences, Massachusetts, USA) at varying cell densities in the range of 0.25–0.75 × 10 5 cells/mL.

Techniques: Concentration Assay

Journal: Scientific Reports

Article Title: Dissecting multi drug resistance in head and neck cancer cells using multicellular tumor spheroids

doi: 10.1038/s41598-019-56273-6

Figure Lengend Snippet: Real-time monitoring of calcein-AM uptake in 2D and MCSs. ( a ) Elucidate calcein-AM uptake in 2D cell cultures and MCSs obtained from LK0917- MCS 17 (upper panel), LK0902- MCS 02 (middle panel), and LK1108- MCS 08 (lower panel) and over a time span of 10 hours with image acquisition at 20-minute intervals (scale bar, 200 μm). ( b ) Heat map pseudo color images of MCSs for differential calcein-AM uptake for the three cell lines (scale bar, 200 μm, scale bar on the right represents the pixel intensity of green fluorescence). ( c ) Mean fluorescence intensity profile with respect to time (12 hours) in the 2D cell cultures (LK0917, LK0902, and LK1108) and MCSs (MCS 17 , MCS 02 , and MCS 08 ) respectively. ( d ) Total accumulated calcein over time (12 hours) in 2D and MCSs for all three cell lines. ( e ) Total accumulated calcein profiles of MCSs obtained from different cell densities. The data are shown as mean ± SD; ***p < 0.001, **p < 0.05, *p = 0.019, and ns = non-significant (n = 3).

Article Snippet: For the generation of MCSs sized 300–500 μm, 200 μL of LK0917 (MCS 17 ), LK0902 (MCS 02 ), and LK1108 (MCS 08 ) single cell suspensions were seeded in ultra-low attachment (ULA) plates (Corning Life Sciences, Massachusetts, USA) at varying cell densities in the range of 0.25–0.75 × 10 5 cells/mL.

Techniques: Fluorescence

Journal: Scientific Reports

Article Title: Dissecting multi drug resistance in head and neck cancer cells using multicellular tumor spheroids

doi: 10.1038/s41598-019-56273-6

Figure Lengend Snippet: ROS activity of all cell lines in 2D and MCSs. ( a ) Live-cell fluorescence images of the 2D (LK0917, LK0902, and LK1108) and MCSs (MCS 17 , MCS 02 , and MCS 08 ) cultures obtained from cell lines in the presence of DCFDA over a period of 60 minutes with image acquisition at 20-minute intervals (upper, middle and lower panel respectively), scale bar, 200 μm). ( b ) Redox state in 2D and MCSs cultures. Data are presented as the mean ± SD; ***p < 0.001, **p < 0.05, *p = 0.019, and no significant differences were observed in case of 2D (n = 3). ( c ) Heat map pseudo color images of MCSs for differential redox status for the three cell lines (scale bar, 200 μm).

Article Snippet: For the generation of MCSs sized 300–500 μm, 200 μL of LK0917 (MCS 17 ), LK0902 (MCS 02 ), and LK1108 (MCS 08 ) single cell suspensions were seeded in ultra-low attachment (ULA) plates (Corning Life Sciences, Massachusetts, USA) at varying cell densities in the range of 0.25–0.75 × 10 5 cells/mL.

Techniques: Activity Assay, Fluorescence

Journal: Scientific Reports

Article Title: Dissecting multi drug resistance in head and neck cancer cells using multicellular tumor spheroids

doi: 10.1038/s41598-019-56273-6

Figure Lengend Snippet: ( a –c) Median fluorescence intensity (MFI) values for 2D and MCSs cultures of LK0917, LK0902, and LK1108 in presence of different ABC pump inhibitors for MDR1, MRP1, and BCRP. ( d ) MFI for MDR1 pump inhibitor (verapamil) and its corresponding live cell imaging for 2D and MCSs cultures. ( e ) Total calcein-AM uptake for the entire course of 10 hours +/− verapamil for 2D and MCSs. The data are shown as a mean of ± SD, ***p < 0.001, **p < 0.05. ( f ) Elucidate calcein-AM uptake in 2D for LK0917, LK0902 and LK1108 in presence and absence of verapamil over a time span of 10 hours with images taken after every 20 minutes (images shown here are at 2 h interval). Scale bar 200 μm.

Article Snippet: For the generation of MCSs sized 300–500 μm, 200 μL of LK0917 (MCS 17 ), LK0902 (MCS 02 ), and LK1108 (MCS 08 ) single cell suspensions were seeded in ultra-low attachment (ULA) plates (Corning Life Sciences, Massachusetts, USA) at varying cell densities in the range of 0.25–0.75 × 10 5 cells/mL.

Techniques: Fluorescence, Live Cell Imaging

Journal: Journal of Microbiology and Biotechnology

Article Title: Deinococcus radiodurans R1 Lysate Induces Tolerogenic Maturation in Lipopolysaccharide-Stimulated Dendritic Cells and Protects Dextran Sulfate Sodium-Induced Colitis in Mice

doi: 10.4014/jmb.2203.03008

Figure Lengend Snippet: Cells were treated with anti-IL-10 mAb (5 ng/ml) or rat IgG (isotype control) for 2 h prior to LPS and DeinoLys treatment. ( A ) Extracellular cytokine levels in the cell culture supernatant were measured using commercial ELISA kits. ( B ) Surface molecules were measured by flow cytometry. The mean fluorescence intensity of each surface molecule in CD11c + cells is represented by bar graphs. ( C ) CD4 + and CD8 + T cells isolated from splenocytes of BALB/c mice were stained with CellTrace™ Violet Cell Proliferation Kit and cocultured with DCs treated with PBS (Con), DeinoLys, LPS, or LPS with DeinoLys in the presence or absence of anti-IL-10 mAb or rat IgG. The proliferation of CD4 + or CD8 + T cells was analyzed by flow cytometry. The mean SD ( n = 3) is shown in all bar graphs. One-way ANOVA was used for statistics, followed by Tukey's multiple comparison. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: Following the established methodology, single-cell suspensions of splenocytes were obtained from 7-week-old BALB/c mice (Orient Bio, Inc.) [ ].

Techniques: Control, Cell Culture, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Fluorescence, Isolation, Staining, Comparison