Journal: RSC Advances

Article Title: CellTrap: an instrument-free microfluidic platform for cell–cell interactions at stochastically generated effector-to-target ratios

doi: 10.1039/d6ra02345b

Figure Lengend Snippet: Characterization of the CellTrap device with 1024 traps. (A) The experimental loading frequencies ( f ) of individual particles, i.e. , green and red, are compared to the theoretical Poisson distributions ( P ) for λ 1 = 0.51 (green) and λ 2 = 0.54 (red). (B) A combinatorial loading distribution of the two particle types was analyzed using a double Poisson distribution, which is compared with the experimental dual loading frequency. (C) Representative bright-field, fluorescence, and overlay images of red and green particles trapped in a channel at different k 1 : k 2 ratios. (D) The experimental loading frequencies ( f ) and the theoretical Poisson distributions ( P ) are plotted for the U87 GFP and NK92 IL2 cells seeded in the CellTrap device with λ 1 = 0.83 (U87 GFP ) and λ 2 = 0.58 (NK92 IL2 ). (E) The experimental dual loading frequency and theoretical double Poisson distributions result in varying E : T or k 1 : k 2 ratios. (F) Representative bright-field, fluorescence, and overlay images of cancer (purple) and immune (white arrows) cells trapped inside a channel at different E : T ratios. (scale bars: 30 µm).

Article Snippet: The human cell lines NK92 IL2 , U87, K562, and LS174T were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Fluorescence

Journal: RSC Advances

Article Title: CellTrap: an instrument-free microfluidic platform for cell–cell interactions at stochastically generated effector-to-target ratios

doi: 10.1039/d6ra02345b

Figure Lengend Snippet: Response of PBMCs and NK92 IL2 against U87 GFP cells. (A) Fluorescence intensity of U87 GFP cells decreases significantly after 4 h of co-incubation with PBMCs at E : T = ≥1 : ≥1. This data is curated from two independent CellTrap devices ( N = 2), where, in total, 97 traps were analyzed ( n = 97). (B) Inside one of the CellTrap devices used in (A), control traps with only one U87 GFP cell per trap, i.e. , E : T = 0 : 1, were analyzed, maintaining a stable fluorescence signal over 14 h ( N = 1, n = 18). (C) Representative time-lapse images of U87 GFP cells interacting with PBMCs at different E : T ratios, along with the control group containing only U87 GFP cells. (D) Fluorescence intensity of U87 GFP cells decreases significantly after 4 h of co-incubation with NK92 IL2 at E : T = 1 : 1. This data is curated from four independent CellTrap devices ( N = 4), where, in total, 213 traps with E : T = 1 : 1 were analyzed ( n = 213). (E) Inside one of the CellTrap devices used in (D), control traps with only one U87 GFP cell per trap, i.e. , E : T = 0 : 1, were analyzed, maintaining a fluorescence signal over 14 h ( N = 1, n = 50). (F) Representative time-lapse images of U87 GFP cells interacting with NK92 IL2 at different E : T ratios, along with the control group containing only U87 GFP cells. (G) Fluorescence intensity of U87 GFP at 0 h and 14 h of co-incubation with NK92 IL2 at E : T = 1 : 1, 1 : 2, 2 : 1 and 2 : 2. The intensity drop is significant across all E : T ratios except 1 : 2. This data is curated from the same CellTrap devices used in (D).

Article Snippet: The human cell lines NK92 IL2 , U87, K562, and LS174T were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Fluorescence, Incubation, Control

Journal: RSC Advances

Article Title: CellTrap: an instrument-free microfluidic platform for cell–cell interactions at stochastically generated effector-to-target ratios

doi: 10.1039/d6ra02345b

Figure Lengend Snippet: Calcium flux and killing response of immune cells against cancer cells. (A) Calcium flux (normalized intensity) in NK92 IL2 immune cells varies over time in the presence of various cancer cell lines (U87, LS174T, K562). NK92 IL2 cells alone show a flat response (control). Each grey line represents a single immune cell tracked. Representative images show NK92 IL2 cells (green) and cancer cells (U87, LS174T, K562) co-incubated in the CellTrap chip at an E : T ratio of 1 : 1. n = number of traps analyzed. (B) The killing response of NK92 IL2 cells at different E : T ratios (1 : 1, 1 : 2, 2 : 1) against cancer cells (U87, LS174T, K562) is quantified and compared with control groups with E : T ratios of ≥1 : 0 and 0 : ≥1. N = number of CellTrap chips analyzed. n = number of traps analyzed. Representative images show NK92 IL2 cells interacting with cancer cells (U87, LS174T, K562 in blue) with varying E : T ratios at 0 and 14 hours. Red color indicates cell death at 14 hours. Scale bars: 25 µm.

Article Snippet: The human cell lines NK92 IL2 , U87, K562, and LS174T were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Control, Incubation

Journal: bioRxiv

Article Title: B7-H3 Modulates Cell Adhesion and Immune Evasion to Promote Tumor Progression and Natural Killer Cell Resistance in Hepatocellular Carcinoma

doi: 10.64898/2026.03.28.714951

Figure Lengend Snippet: (A) Flow cytometric analysis of PD-L1, PD-L2, CD47, and CD24 expression in B7-H3–knockdown SNU449 cells. (B) Quantification of MFI shown in (A), expressed as relative MFI (%) compared with siCon cells. (C) Flow cytometric analysis of PD-L1, PD-L2, and CD47 expression in control and B7-H3 KO SNU449 cells. (D) Quantification of MFI shown in (C), expressed as relative MFI (%) compared with control cells. (E) Western blot analysis of immune checkpoint proteins in control and B7-H3 KO SNU449 cells. Protein levels were quantified using ImageJ software with GAPDH as a loading control. (F) NK92 cell–mediated cytotoxicity assay. Control and B7-H3 KO SNU449 cells were co-cultured with NK92 cells at a 1:1 effector-to-target (E:T) ratio for 48 hours, followed by crystal violet staining of surviving tumor cells. Representative images from three independent experiments are shown. (G) Quantification of surviving cells shown in (F) by measuring absorbance at 590 nm. (H) NK92 cell–mediated cytotoxicity assessed by calcein-AM release assay at various E:T ratios. (I) Primary NK cell–mediated cytotoxicity assessed by calcein-AM release assay at various E:T ratios. ns, not significant; *, p < 0.05; **, p < 0.01; ***, p < 0.005.

Article Snippet: The NK92 cell line was purchased from ATCC and cultured in α-MEM (Biowest) medium supplemented with 12.5% FBS (Biowest), 12.5% horse serum (Biowest), 200μM inositol (Sigma-Aldrich, St.Lois, MO, USA), 20μM Folic acid (Sigma-Aldrich), 100μM 2-mercaptoethanol (Sigma-Aldrich), AA solution (Biowest), and 400 unit/ml IL2 (Peprotech, Rocky Hill, NJ, USA).

Techniques: Expressing, Knockdown, Control, Western Blot, Software, Cytotoxicity Assay, Cell Culture, Staining, Release Assay