Journal: PLoS ONE

Article Title: Anti-Tumor Effect against Human Cancer Xenografts by a Fully Human Monoclonal Antibody to a Variant 8-Epitope of CD44R1 Expressed on Cancer Stem Cells

doi: 10.1371/journal.pone.0029728

Figure Lengend Snippet: GV5, Cetuximab and Trastuzumab were compared for the reactivity with HUVEC, NHEK and HCT116 by FCM (histograms) with an Accuri C6 flow cytometer (A). Reactivity of GV5 with various human cells was analyzed by FCM, and depicted as histograms (B) using a BD-LSR flow cytometer and as ΔMFI (C) using an Accuri C6 flow cytometer.

Article Snippet: After three washes with PBS, cells were suspended in PBS containing 0.2% BSA, and analyzed with an Accuri C6 flow cytometer (Tomy Digital Biology, Tokyo, Japan) or a BD-LSR flow cytometer (Becton-Dickinson, Sunnyvale, CA, USA).

Techniques: Flow Cytometry

Journal: PLoS ONE

Article Title: Anti-Tumor Effect against Human Cancer Xenografts by a Fully Human Monoclonal Antibody to a Variant 8-Epitope of CD44R1 Expressed on Cancer Stem Cells

doi: 10.1371/journal.pone.0029728

Figure Lengend Snippet: (A) HSC3 cells were cultured with or without GV5 (10 µg/ml) for 12 h, and were immunostained with GV5 followed by FITC-conjugated donkey anti-human IgG (H+L). Cell-surface CD44R1 proteins in these cells were detected by FCM. Experiments were repeated three times, and similar results were obtained. (B) After HSC-3 cells and sera for complement and mAb were mixed and incubated in each well of U-bottomed 96-well plate for 1 h, DAPI was added to each well. Percentages of DAPI-stained cells (dead cells) were calculated by FCM. Experiments were repeated three times, and similar results were obtained. (C) HSC-3 cells (1.5×10 5 ) were mixed with effector cells of splenocytes (3×10 6 or 9×10 6 ) from KNS nude mice, which were pre-cultured overnight with lL-2, with or without mAb, in each well of U-bottomed 96-well plate for 5 h, and PI was added to each well. Cytotoxicity was analyzed using an Accuri flow cytometer. R1 , HSC3 human target cells; R2 , mouse effector cells; R3 , PI-stained dead cells in R1 ; R4 , PI-unstained living cells in R1 . (D) Cell death (%) of HSC-3 cells by mouse effector cells (E/T = 0, 20 or 60) with or without GV5 was plotted. E/T means effector to target ratio.

Article Snippet: After three washes with PBS, cells were suspended in PBS containing 0.2% BSA, and analyzed with an Accuri C6 flow cytometer (Tomy Digital Biology, Tokyo, Japan) or a BD-LSR flow cytometer (Becton-Dickinson, Sunnyvale, CA, USA).

Techniques: Cell Culture, Incubation, Staining, Flow Cytometry