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ATCC natural killer cell line nk 92mi

Microphysiological system “NNTI-chip” models the preferential migration of natural killer (NK) cells toward different neutrophil subtypes in scenario 1. (A) Side channels A and B of the NNTI-chip housed two different human neutrophil subtypes (i.e., N1 and N2 neutrophils, N1 and N0 neutrophils, or N2 and N0 neutrophils, respectively) acting as two competitive signals, and NK cells in the central channel were allowed to migrate preferentially into either side channel A or B. Both neutrophils and NK cells were embedded in a 3D collagen hydrogel to mimic the extracellular matrix of the tumor tissue. Created with Rhino 7 and BioRender.com. (B) Representative 10X bright-field and epifluorescence images showing N1 neutrophils (orange) in side channel A, N2 neutrophils (magenta) in side channel B, and NK cells (blue) in the central channel immediately after loading of NK cells into the NNTI-chip. The three channels are separated and interconnected by PDMS microposts. Scale bar, 100 μm. (C) The workflow of a typical experiment for scenario 1. <t>NK</t> <t>cell</t> migration toward neutrophils in both side channels and NK cell motility after migration into side channels were quantified as readouts. Created with BioRender.com.

Natural Killer Cell Line Nk 92mi, supplied by ATCC, used in various techniques. Bioz Stars score: 96/100, based on 418 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 96 stars, based on 418 article reviews

natural killer cell line nk 92mi - by Bioz Stars, 2026-08

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1) Product Images from "Quantifying the Dual Effect of Antitumor and Pro-Tumor Human Neutrophils on Natural Killer Cell Behaviors in a Microphysiological System"

Article Title: Quantifying the Dual Effect of Antitumor and Pro-Tumor Human Neutrophils on Natural Killer Cell Behaviors in a Microphysiological System

Journal: ACS Biomaterials Science & Engineering

doi: 10.1021/acsbiomaterials.5c02083

Figure Legend Snippet: Microphysiological system “NNTI-chip” models the preferential migration of natural killer (NK) cells toward different neutrophil subtypes in scenario 1. (A) Side channels A and B of the NNTI-chip housed two different human neutrophil subtypes (i.e., N1 and N2 neutrophils, N1 and N0 neutrophils, or N2 and N0 neutrophils, respectively) acting as two competitive signals, and NK cells in the central channel were allowed to migrate preferentially into either side channel A or B. Both neutrophils and NK cells were embedded in a 3D collagen hydrogel to mimic the extracellular matrix of the tumor tissue. Created with Rhino 7 and BioRender.com. (B) Representative 10X bright-field and epifluorescence images showing N1 neutrophils (orange) in side channel A, N2 neutrophils (magenta) in side channel B, and NK cells (blue) in the central channel immediately after loading of NK cells into the NNTI-chip. The three channels are separated and interconnected by PDMS microposts. Scale bar, 100 μm. (C) The workflow of a typical experiment for scenario 1. NK cell migration toward neutrophils in both side channels and NK cell motility after migration into side channels were quantified as readouts. Created with BioRender.com.

Techniques Used: Migration

NK-92MI cells show preferential migration toward N1-like neutrophils over N2-like neutrophils. (A) Representative 10X images showing NK-92MI cells (white) migrating from the central channel of the NNTI-chip into side channels A and B under three competitive conditions: N1-like vs N2-like neutrophils, N1-like vs N0 neutrophils, and N2-like vs N0 neutrophils (unstained) at t = 0, 12, and 24 h as representative time points. The yellow dashed line marks the boundary between the central channel and the side channels. Scale bar, 100 μm. (B) (i) Line graphs showing the percentage of NK cell migration, defined as the number of NK-92MI cells in side channel A or B at a given time point divided by the initial number of NK-92MI cells in the central channel at t = 0 h, every 2 h over 24 h in each competitive condition. Bars show the mean ± SD; n = 8–9 chips per condition. Statistical significance is shown for t = 24 h. (ii) Line graphs showing the rate of NK cell migration, defined as the increase in the percentage of NK cell migration per hour, every 2 h over 24 h under each competitive condition. Bars show mean ± SD. (C) The ratio of NK cell migration in side channel B over side channel A at t = 24 h in three competitive conditions: N1/N2, N1/N0, and N2/N0. Each data point represents an NNTI-chip and n = 8–9 chips per condition. (D) The maximum rate of NK cell migration toward N0, N1-like, and N2-like neutrophils, defined as the highest rate of NK cell migration at any time point over 24 h. Each data point represents a side channel and n = 16–18 side channels per condition. Bars show mean ± SD with the mean values written above the points. Five independent experiments were performed. ns: p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, paired t test in (Bi); ANOVA with Tukey multiple comparisons test in (C) and (D).

Figure Legend Snippet: NK-92MI cells show preferential migration toward N1-like neutrophils over N2-like neutrophils. (A) Representative 10X images showing NK-92MI cells (white) migrating from the central channel of the NNTI-chip into side channels A and B under three competitive conditions: N1-like vs N2-like neutrophils, N1-like vs N0 neutrophils, and N2-like vs N0 neutrophils (unstained) at t = 0, 12, and 24 h as representative time points. The yellow dashed line marks the boundary between the central channel and the side channels. Scale bar, 100 μm. (B) (i) Line graphs showing the percentage of NK cell migration, defined as the number of NK-92MI cells in side channel A or B at a given time point divided by the initial number of NK-92MI cells in the central channel at t = 0 h, every 2 h over 24 h in each competitive condition. Bars show the mean ± SD; n = 8–9 chips per condition. Statistical significance is shown for t = 24 h. (ii) Line graphs showing the rate of NK cell migration, defined as the increase in the percentage of NK cell migration per hour, every 2 h over 24 h under each competitive condition. Bars show mean ± SD. (C) The ratio of NK cell migration in side channel B over side channel A at t = 24 h in three competitive conditions: N1/N2, N1/N0, and N2/N0. Each data point represents an NNTI-chip and n = 8–9 chips per condition. (D) The maximum rate of NK cell migration toward N0, N1-like, and N2-like neutrophils, defined as the highest rate of NK cell migration at any time point over 24 h. Each data point represents a side channel and n = 16–18 side channels per condition. Bars show mean ± SD with the mean values written above the points. Five independent experiments were performed. ns: p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, paired t test in (Bi); ANOVA with Tukey multiple comparisons test in (C) and (D).

Techniques Used: Migration

NK-92MI cells show lower motility after migration toward N1-like neutrophils than N2-like neutrophils. (A) Representative 10X images showing the single-cell trajectories (color-coded) of NK-92MI cells (white) after migration into side channel A or B housing neutrophils (not shown) at times of 6, 12, 18, and 24 h of time-lapse imaging. NK-92MI cells were tracked over 30 min intervals every 6 h using TrackMate (ImageJ). Scale bar, 100 μm. (B) Definitions and significance of the following motility parameters of a single-cell trajectory: speed, displacement, and directionality. (C) Line graphs showing the speed (i), displacement (ii), and directionality (iii) of motile NK-92MI cells after migration toward N0, N1-like, and N2-like neutrophils over time at t = 6, 12, 18, and 24 h. The median with 25% and 75% percentiles as error bars is shown. n = 597–882 cells per condition. Individual data points were not shown for clean data visualization. Asterisks (*) represent the significance of differences between N1 and N2 conditions at each time point, whereas pound signs (#) between t = 6 h and t = 24 h for each condition (N0, N1, or N2). ns: p ≥ 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001, Kruskal–Wallis test. (D) Violin plots showing the speed (i), displacement (ii), and directionality (iii) of motile NK-92MI cells with single-cell resolution at t = 12 h as a representative time point in specified conditions. Each data point represents a single NK-92MI cell and n = 597–882 cells from 17 to 18 side channels of the NNTI-chips per condition. The yellow solid lines show the medians, and the yellow dashed lines show 25% and 75% percentiles. Median values are written above the points. Five independent experiments were performed. ns: p ≥ 0.05, * p < 0.05, **** p < 0.0001, Kruskal–Wallis test.

Figure Legend Snippet: NK-92MI cells show lower motility after migration toward N1-like neutrophils than N2-like neutrophils. (A) Representative 10X images showing the single-cell trajectories (color-coded) of NK-92MI cells (white) after migration into side channel A or B housing neutrophils (not shown) at times of 6, 12, 18, and 24 h of time-lapse imaging. NK-92MI cells were tracked over 30 min intervals every 6 h using TrackMate (ImageJ). Scale bar, 100 μm. (B) Definitions and significance of the following motility parameters of a single-cell trajectory: speed, displacement, and directionality. (C) Line graphs showing the speed (i), displacement (ii), and directionality (iii) of motile NK-92MI cells after migration toward N0, N1-like, and N2-like neutrophils over time at t = 6, 12, 18, and 24 h. The median with 25% and 75% percentiles as error bars is shown. n = 597–882 cells per condition. Individual data points were not shown for clean data visualization. Asterisks (*) represent the significance of differences between N1 and N2 conditions at each time point, whereas pound signs (#) between t = 6 h and t = 24 h for each condition (N0, N1, or N2). ns: p ≥ 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001, Kruskal–Wallis test. (D) Violin plots showing the speed (i), displacement (ii), and directionality (iii) of motile NK-92MI cells with single-cell resolution at t = 12 h as a representative time point in specified conditions. Each data point represents a single NK-92MI cell and n = 597–882 cells from 17 to 18 side channels of the NNTI-chips per condition. The yellow solid lines show the medians, and the yellow dashed lines show 25% and 75% percentiles. Median values are written above the points. Five independent experiments were performed. ns: p ≥ 0.05, * p < 0.05, **** p < 0.0001, Kruskal–Wallis test.

Techniques Used: Migration, Single Cell, Imaging

Microphysiological system “NNTI-chip” models the tumor cytotoxicity and infiltration of NK cells in the presence of different neutrophil subtypes in scenario 2. (A) A mixture of tumor spheroids, NK cells, and neutrophils (N0, N1, or N2) was embedded in a 3D collagen hydrogel and seeded in the side channels of the NNTI-chip, which mimics the solid tumor tissue. Side channels A and B served as technical replicates. Created with Rhino 7 and BioRender.com. (B) Representative 10X brightfield and epifluorescence images showing tumor spheroids (red), NK cells (cyan), and neutrophils (magenta) in the same side channel of the NNTI-chip. Scale bar, 100 μm. (C) (i) The frequency distribution of diameters of tumor spheroids loaded into the two side channels of the NNTI-chip. The mean diameter is 93.0 ± 18.8 μm ( n = 873 spheroids). (ii) The frequency distribution of the number of tumor spheroids loaded into each side channel of the NNTI-chip. The mean number is 11.3 ± 4.0 ( n = 75 side channels). (D) The workflow of a typical experiment for scenario 2. Real-time tumor spheroid apoptosis in 2D, end-point tumor spheroid apoptosis in 3D, and NK cell infiltration in tumor spheroids were quantified as readouts. Created with BioRender.com.

Figure Legend Snippet: Microphysiological system “NNTI-chip” models the tumor cytotoxicity and infiltration of NK cells in the presence of different neutrophil subtypes in scenario 2. (A) A mixture of tumor spheroids, NK cells, and neutrophils (N0, N1, or N2) was embedded in a 3D collagen hydrogel and seeded in the side channels of the NNTI-chip, which mimics the solid tumor tissue. Side channels A and B served as technical replicates. Created with Rhino 7 and BioRender.com. (B) Representative 10X brightfield and epifluorescence images showing tumor spheroids (red), NK cells (cyan), and neutrophils (magenta) in the same side channel of the NNTI-chip. Scale bar, 100 μm. (C) (i) The frequency distribution of diameters of tumor spheroids loaded into the two side channels of the NNTI-chip. The mean diameter is 93.0 ± 18.8 μm ( n = 873 spheroids). (ii) The frequency distribution of the number of tumor spheroids loaded into each side channel of the NNTI-chip. The mean number is 11.3 ± 4.0 ( n = 75 side channels). (D) The workflow of a typical experiment for scenario 2. Real-time tumor spheroid apoptosis in 2D, end-point tumor spheroid apoptosis in 3D, and NK cell infiltration in tumor spheroids were quantified as readouts. Created with BioRender.com.

Techniques Used:

N1-like neutrophils restore the cytotoxicity of NK-92MI cells against PANC-1 tumor spheroids, while N2-like neutrophils suppress it. (A) Representative 10X epifluorescence images showing the apoptosis (caspase-3/7 green) of tumor spheroids (red) in the NNTI-chip at 0, 6, 12, 18, and 24 h as representative time points in the following conditions: alone (control), with NK-92MI cells (blue), with NK-92MI cells and N0, N1-like, or N2-like neutrophils (unstained). Scale bar, 50 μm. (B) (i) Line graph showing the temporal dynamics of tumor spheroid apoptosis, quantified as normalized caspase-3/7 green intensity per spheroid, every 2 h over 24 h in all five conditions. Bars show mean ± SD. (ii) Bar plot showing tumor spheroid apoptosis at t = 24 h. Bars show mean ± SD with mean values written above the points. Each data point represents a tumor spheroid and n = 68–83 spheroids per condition. (C) (i) Line graph showing the rate of tumor apoptosis, defined as the increase in tumor spheroid apoptosis per hour, every 2 h over 24 h in all five conditions. Bars show mean ± SD. (ii) Bar plot showing the maximum rate of tumor apoptosis, defined as the highest rate of tumor apoptosis at any time point over 24 h. Bars show mean ± SD. (D) 3D rendering of representative 10X confocal images showing the apoptosis (caspase-3/7 green) of tumor spheroids (red) fixed at t = 24 h and the extraction of the apoptotic volume and the total spheroid volume by Imaris. Images were acquired as z-stacks with a step size of 2 μm. Apoptotic signals colocalized with DiD-stained NK-92MI cells were excluded to remove apoptotic NK cells. Tumor apoptosis was quantified as the apoptotic volume divided by the total spheroid volume. Scale bar: 30 μm. (E) Bar plot showing the percentage of apoptotic volume per tumor spheroid. Each data point represents a spheroid and n = 58–72 spheroids per condition. Bars show mean ± SD with mean values written above the points. At least three independent experiments were performed. ns: p ≥ 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, Kruskal–Wallis test.

Figure Legend Snippet: N1-like neutrophils restore the cytotoxicity of NK-92MI cells against PANC-1 tumor spheroids, while N2-like neutrophils suppress it. (A) Representative 10X epifluorescence images showing the apoptosis (caspase-3/7 green) of tumor spheroids (red) in the NNTI-chip at 0, 6, 12, 18, and 24 h as representative time points in the following conditions: alone (control), with NK-92MI cells (blue), with NK-92MI cells and N0, N1-like, or N2-like neutrophils (unstained). Scale bar, 50 μm. (B) (i) Line graph showing the temporal dynamics of tumor spheroid apoptosis, quantified as normalized caspase-3/7 green intensity per spheroid, every 2 h over 24 h in all five conditions. Bars show mean ± SD. (ii) Bar plot showing tumor spheroid apoptosis at t = 24 h. Bars show mean ± SD with mean values written above the points. Each data point represents a tumor spheroid and n = 68–83 spheroids per condition. (C) (i) Line graph showing the rate of tumor apoptosis, defined as the increase in tumor spheroid apoptosis per hour, every 2 h over 24 h in all five conditions. Bars show mean ± SD. (ii) Bar plot showing the maximum rate of tumor apoptosis, defined as the highest rate of tumor apoptosis at any time point over 24 h. Bars show mean ± SD. (D) 3D rendering of representative 10X confocal images showing the apoptosis (caspase-3/7 green) of tumor spheroids (red) fixed at t = 24 h and the extraction of the apoptotic volume and the total spheroid volume by Imaris. Images were acquired as z-stacks with a step size of 2 μm. Apoptotic signals colocalized with DiD-stained NK-92MI cells were excluded to remove apoptotic NK cells. Tumor apoptosis was quantified as the apoptotic volume divided by the total spheroid volume. Scale bar: 30 μm. (E) Bar plot showing the percentage of apoptotic volume per tumor spheroid. Each data point represents a spheroid and n = 58–72 spheroids per condition. Bars show mean ± SD with mean values written above the points. At least three independent experiments were performed. ns: p ≥ 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, Kruskal–Wallis test.

Techniques Used: Control, Extraction, Staining

Both N1-like and N2-like neutrophils inhibit the infiltration of NK-92MI cells into PANC-1 tumor spheroids. (A) 3D rendering of representative 10X confocal images showing the infiltration of NK-92MI cells (cyan) in tumor spheroids (red) in the absence (control) or presence of N0, N1-like, or N2-like neutrophils (unstained) in the NNTI-chip fixed at 24 h and the extraction of the volumes of tumor spheroids and infiltrated NK cells by Imaris. Images were acquired as z-stacks with a 2 μm step size. NK cell infiltration was quantified as the volume of infiltrated NK-92MI cells divided by the total spheroid volume. Scale bar, 30 μm. (B) Bar plot showing the percentage of infiltrated NK-92MI cells per tumor spheroid in specified conditions. Each data point represents a spheroid and n = 58–77 spheroids per condition. Bars show mean ± SD with mean values written above the points. At least three independent experiments were performed. ns: p ≥ 0.05, ** p < 0.01, **** p < 0.0001, Kruskal–Wallis test.

Figure Legend Snippet: Both N1-like and N2-like neutrophils inhibit the infiltration of NK-92MI cells into PANC-1 tumor spheroids. (A) 3D rendering of representative 10X confocal images showing the infiltration of NK-92MI cells (cyan) in tumor spheroids (red) in the absence (control) or presence of N0, N1-like, or N2-like neutrophils (unstained) in the NNTI-chip fixed at 24 h and the extraction of the volumes of tumor spheroids and infiltrated NK cells by Imaris. Images were acquired as z-stacks with a 2 μm step size. NK cell infiltration was quantified as the volume of infiltrated NK-92MI cells divided by the total spheroid volume. Scale bar, 30 μm. (B) Bar plot showing the percentage of infiltrated NK-92MI cells per tumor spheroid in specified conditions. Each data point represents a spheroid and n = 58–77 spheroids per condition. Bars show mean ± SD with mean values written above the points. At least three independent experiments were performed. ns: p ≥ 0.05, ** p < 0.01, **** p < 0.0001, Kruskal–Wallis test.

Techniques Used: Control, Extraction

Image Search Results

Journal: ACS Biomaterials Science & Engineering

Article Title: Quantifying the Dual Effect of Antitumor and Pro-Tumor Human Neutrophils on Natural Killer Cell Behaviors in a Microphysiological System

doi: 10.1021/acsbiomaterials.5c02083

Figure Lengend Snippet: Microphysiological system “NNTI-chip” models the preferential migration of natural killer (NK) cells toward different neutrophil subtypes in scenario 1. (A) Side channels A and B of the NNTI-chip housed two different human neutrophil subtypes (i.e., N1 and N2 neutrophils, N1 and N0 neutrophils, or N2 and N0 neutrophils, respectively) acting as two competitive signals, and NK cells in the central channel were allowed to migrate preferentially into either side channel A or B. Both neutrophils and NK cells were embedded in a 3D collagen hydrogel to mimic the extracellular matrix of the tumor tissue. Created with Rhino 7 and BioRender.com. (B) Representative 10X bright-field and epifluorescence images showing N1 neutrophils (orange) in side channel A, N2 neutrophils (magenta) in side channel B, and NK cells (blue) in the central channel immediately after loading of NK cells into the NNTI-chip. The three channels are separated and interconnected by PDMS microposts. Scale bar, 100 μm. (C) The workflow of a typical experiment for scenario 1. NK cell migration toward neutrophils in both side channels and NK cell motility after migration into side channels were quantified as readouts. Created with BioRender.com.

Article Snippet: The human natural killer cell line NK-92MI (American Type Culture Collection ATCC, CRL-2408) was cultured in Roswell Park Memorial Institute (RPMI) 1640 (Corning, 10040CV) supplemented with 10% fetal bovine serum (FBS, Corning, 35015CV), 1% sodium pyruvate (Gibco, 11360070), 1% minimum essential medium (MEM) nonessential amino acids solution (Gibco, 11140050), 1% GlutaMAX (Gibco, 35050061), 0.1% 2-mercaptoethanol (Gibco, 21985023), and 1% penicillin–streptomycin (Cytiva), denoted as complete RPMI.

Techniques: Migration

Journal: ACS Biomaterials Science & Engineering

Article Title: Quantifying the Dual Effect of Antitumor and Pro-Tumor Human Neutrophils on Natural Killer Cell Behaviors in a Microphysiological System

doi: 10.1021/acsbiomaterials.5c02083

Figure Lengend Snippet: NK-92MI cells show preferential migration toward N1-like neutrophils over N2-like neutrophils. (A) Representative 10X images showing NK-92MI cells (white) migrating from the central channel of the NNTI-chip into side channels A and B under three competitive conditions: N1-like vs N2-like neutrophils, N1-like vs N0 neutrophils, and N2-like vs N0 neutrophils (unstained) at t = 0, 12, and 24 h as representative time points. The yellow dashed line marks the boundary between the central channel and the side channels. Scale bar, 100 μm. (B) (i) Line graphs showing the percentage of NK cell migration, defined as the number of NK-92MI cells in side channel A or B at a given time point divided by the initial number of NK-92MI cells in the central channel at t = 0 h, every 2 h over 24 h in each competitive condition. Bars show the mean ± SD; n = 8–9 chips per condition. Statistical significance is shown for t = 24 h. (ii) Line graphs showing the rate of NK cell migration, defined as the increase in the percentage of NK cell migration per hour, every 2 h over 24 h under each competitive condition. Bars show mean ± SD. (C) The ratio of NK cell migration in side channel B over side channel A at t = 24 h in three competitive conditions: N1/N2, N1/N0, and N2/N0. Each data point represents an NNTI-chip and n = 8–9 chips per condition. (D) The maximum rate of NK cell migration toward N0, N1-like, and N2-like neutrophils, defined as the highest rate of NK cell migration at any time point over 24 h. Each data point represents a side channel and n = 16–18 side channels per condition. Bars show mean ± SD with the mean values written above the points. Five independent experiments were performed. ns: p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, paired t test in (Bi); ANOVA with Tukey multiple comparisons test in (C) and (D).

Techniques: Migration

Journal: ACS Biomaterials Science & Engineering

Article Title: Quantifying the Dual Effect of Antitumor and Pro-Tumor Human Neutrophils on Natural Killer Cell Behaviors in a Microphysiological System

doi: 10.1021/acsbiomaterials.5c02083

Figure Lengend Snippet: NK-92MI cells show lower motility after migration toward N1-like neutrophils than N2-like neutrophils. (A) Representative 10X images showing the single-cell trajectories (color-coded) of NK-92MI cells (white) after migration into side channel A or B housing neutrophils (not shown) at times of 6, 12, 18, and 24 h of time-lapse imaging. NK-92MI cells were tracked over 30 min intervals every 6 h using TrackMate (ImageJ). Scale bar, 100 μm. (B) Definitions and significance of the following motility parameters of a single-cell trajectory: speed, displacement, and directionality. (C) Line graphs showing the speed (i), displacement (ii), and directionality (iii) of motile NK-92MI cells after migration toward N0, N1-like, and N2-like neutrophils over time at t = 6, 12, 18, and 24 h. The median with 25% and 75% percentiles as error bars is shown. n = 597–882 cells per condition. Individual data points were not shown for clean data visualization. Asterisks (*) represent the significance of differences between N1 and N2 conditions at each time point, whereas pound signs (#) between t = 6 h and t = 24 h for each condition (N0, N1, or N2). ns: p ≥ 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001, Kruskal–Wallis test. (D) Violin plots showing the speed (i), displacement (ii), and directionality (iii) of motile NK-92MI cells with single-cell resolution at t = 12 h as a representative time point in specified conditions. Each data point represents a single NK-92MI cell and n = 597–882 cells from 17 to 18 side channels of the NNTI-chips per condition. The yellow solid lines show the medians, and the yellow dashed lines show 25% and 75% percentiles. Median values are written above the points. Five independent experiments were performed. ns: p ≥ 0.05, * p < 0.05, **** p < 0.0001, Kruskal–Wallis test.

Techniques: Migration, Single Cell, Imaging

Journal: ACS Biomaterials Science & Engineering

Article Title: Quantifying the Dual Effect of Antitumor and Pro-Tumor Human Neutrophils on Natural Killer Cell Behaviors in a Microphysiological System

doi: 10.1021/acsbiomaterials.5c02083

Figure Lengend Snippet: Microphysiological system “NNTI-chip” models the tumor cytotoxicity and infiltration of NK cells in the presence of different neutrophil subtypes in scenario 2. (A) A mixture of tumor spheroids, NK cells, and neutrophils (N0, N1, or N2) was embedded in a 3D collagen hydrogel and seeded in the side channels of the NNTI-chip, which mimics the solid tumor tissue. Side channels A and B served as technical replicates. Created with Rhino 7 and BioRender.com. (B) Representative 10X brightfield and epifluorescence images showing tumor spheroids (red), NK cells (cyan), and neutrophils (magenta) in the same side channel of the NNTI-chip. Scale bar, 100 μm. (C) (i) The frequency distribution of diameters of tumor spheroids loaded into the two side channels of the NNTI-chip. The mean diameter is 93.0 ± 18.8 μm ( n = 873 spheroids). (ii) The frequency distribution of the number of tumor spheroids loaded into each side channel of the NNTI-chip. The mean number is 11.3 ± 4.0 ( n = 75 side channels). (D) The workflow of a typical experiment for scenario 2. Real-time tumor spheroid apoptosis in 2D, end-point tumor spheroid apoptosis in 3D, and NK cell infiltration in tumor spheroids were quantified as readouts. Created with BioRender.com.

Techniques:

Journal: ACS Biomaterials Science & Engineering

Article Title: Quantifying the Dual Effect of Antitumor and Pro-Tumor Human Neutrophils on Natural Killer Cell Behaviors in a Microphysiological System

doi: 10.1021/acsbiomaterials.5c02083

Figure Lengend Snippet: N1-like neutrophils restore the cytotoxicity of NK-92MI cells against PANC-1 tumor spheroids, while N2-like neutrophils suppress it. (A) Representative 10X epifluorescence images showing the apoptosis (caspase-3/7 green) of tumor spheroids (red) in the NNTI-chip at 0, 6, 12, 18, and 24 h as representative time points in the following conditions: alone (control), with NK-92MI cells (blue), with NK-92MI cells and N0, N1-like, or N2-like neutrophils (unstained). Scale bar, 50 μm. (B) (i) Line graph showing the temporal dynamics of tumor spheroid apoptosis, quantified as normalized caspase-3/7 green intensity per spheroid, every 2 h over 24 h in all five conditions. Bars show mean ± SD. (ii) Bar plot showing tumor spheroid apoptosis at t = 24 h. Bars show mean ± SD with mean values written above the points. Each data point represents a tumor spheroid and n = 68–83 spheroids per condition. (C) (i) Line graph showing the rate of tumor apoptosis, defined as the increase in tumor spheroid apoptosis per hour, every 2 h over 24 h in all five conditions. Bars show mean ± SD. (ii) Bar plot showing the maximum rate of tumor apoptosis, defined as the highest rate of tumor apoptosis at any time point over 24 h. Bars show mean ± SD. (D) 3D rendering of representative 10X confocal images showing the apoptosis (caspase-3/7 green) of tumor spheroids (red) fixed at t = 24 h and the extraction of the apoptotic volume and the total spheroid volume by Imaris. Images were acquired as z-stacks with a step size of 2 μm. Apoptotic signals colocalized with DiD-stained NK-92MI cells were excluded to remove apoptotic NK cells. Tumor apoptosis was quantified as the apoptotic volume divided by the total spheroid volume. Scale bar: 30 μm. (E) Bar plot showing the percentage of apoptotic volume per tumor spheroid. Each data point represents a spheroid and n = 58–72 spheroids per condition. Bars show mean ± SD with mean values written above the points. At least three independent experiments were performed. ns: p ≥ 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, Kruskal–Wallis test.

Techniques: Control, Extraction, Staining

Journal: ACS Biomaterials Science & Engineering

Article Title: Quantifying the Dual Effect of Antitumor and Pro-Tumor Human Neutrophils on Natural Killer Cell Behaviors in a Microphysiological System

doi: 10.1021/acsbiomaterials.5c02083

Figure Lengend Snippet: Both N1-like and N2-like neutrophils inhibit the infiltration of NK-92MI cells into PANC-1 tumor spheroids. (A) 3D rendering of representative 10X confocal images showing the infiltration of NK-92MI cells (cyan) in tumor spheroids (red) in the absence (control) or presence of N0, N1-like, or N2-like neutrophils (unstained) in the NNTI-chip fixed at 24 h and the extraction of the volumes of tumor spheroids and infiltrated NK cells by Imaris. Images were acquired as z-stacks with a 2 μm step size. NK cell infiltration was quantified as the volume of infiltrated NK-92MI cells divided by the total spheroid volume. Scale bar, 30 μm. (B) Bar plot showing the percentage of infiltrated NK-92MI cells per tumor spheroid in specified conditions. Each data point represents a spheroid and n = 58–77 spheroids per condition. Bars show mean ± SD with mean values written above the points. At least three independent experiments were performed. ns: p ≥ 0.05, ** p < 0.01, **** p < 0.0001, Kruskal–Wallis test.

Techniques: Control, Extraction

Poxin protects NK-92 cells from pDNA transfection-induced innate immunity. A. Cells were transfected by electroporation with 10 μg pDNA, and mRNA dynamics were analyzed by RT-qPCR (n = 3). B. IFN-β secretion levels were quantitatively analyzed using ELISA. C. Expression levels of innate immune signaling proteins were detected by western blotting. (Representative data shown). D. Quantitative analysis of p-STING and p-TBK1 expression levels.

Journal: Journal of Advanced Research

Article Title: Enhancement of NK cell activity via DNA-sensing inhibition by Poxin transgene

doi: 10.1016/j.jare.2025.05.058

Figure Lengend Snippet: Poxin protects NK-92 cells from pDNA transfection-induced innate immunity. A. Cells were transfected by electroporation with 10 μg pDNA, and mRNA dynamics were analyzed by RT-qPCR (n = 3). B. IFN-β secretion levels were quantitatively analyzed using ELISA. C. Expression levels of innate immune signaling proteins were detected by western blotting. (Representative data shown). D. Quantitative analysis of p-STING and p-TBK1 expression levels.

Article Snippet: The human natural killer cell line NK-92 (CRL-2407) and human monocyte cell line THP-1 (TIB-202) were obtained from ATCC and cultured at 37 °C in an atmosphere of 95 % air and 5 % CO 2 .

Techniques: Transfection, Electroporation, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot

Irradiation-induced apoptosis and DNA sensing in NK-92 cells through endogenous dsDNA. A. Cell survival rate was measured by FACS using an apoptosis detection kit at specified time points after irradiation. B. Cell proliferation rate was quantified by counting live cells. C. Cytotoxicity of NK-92 cells against K562 cells over 4 h was assessed. (Significance: a = * p < 0.05; b = ** p < 0.01; c = *** p < 0.001; d = **** p < 0.0001). D. Protein expression levels were evaluated by western blotting. (Representative data shown). Quantitative analysis of p-TBK1 expression level is shown.

Journal: Journal of Advanced Research

Article Title: Enhancement of NK cell activity via DNA-sensing inhibition by Poxin transgene

doi: 10.1016/j.jare.2025.05.058

Figure Lengend Snippet: Irradiation-induced apoptosis and DNA sensing in NK-92 cells through endogenous dsDNA. A. Cell survival rate was measured by FACS using an apoptosis detection kit at specified time points after irradiation. B. Cell proliferation rate was quantified by counting live cells. C. Cytotoxicity of NK-92 cells against K562 cells over 4 h was assessed. (Significance: a = * p < 0.05; b = ** p < 0.01; c = *** p < 0.001; d = **** p < 0.0001). D. Protein expression levels were evaluated by western blotting. (Representative data shown). Quantitative analysis of p-TBK1 expression level is shown.

Techniques: Irradiation, Expressing, Western Blot

Poxin prolongs NK-92 cell survival and activity by reducing irradiation-induced innate immune activation. A. Cell survival rate was measured by FACS at specified times after irradiation. B. Cell proliferation rate was quantified by counting live cells. C. Cytotoxicity of effector cells against K562 cells was measured 1- and 2-days post-irradiation over a 4-hour period. D. Protein expression levels were detected by western blotting. (Representative data shown). E. Quantitative analysis of p-IRF3 expression level.

Journal: Journal of Advanced Research

Article Title: Enhancement of NK cell activity via DNA-sensing inhibition by Poxin transgene

doi: 10.1016/j.jare.2025.05.058

Figure Lengend Snippet: Poxin prolongs NK-92 cell survival and activity by reducing irradiation-induced innate immune activation. A. Cell survival rate was measured by FACS at specified times after irradiation. B. Cell proliferation rate was quantified by counting live cells. C. Cytotoxicity of effector cells against K562 cells was measured 1- and 2-days post-irradiation over a 4-hour period. D. Protein expression levels were detected by western blotting. (Representative data shown). E. Quantitative analysis of p-IRF3 expression level.

Techniques: Activity Assay, Irradiation, Activation Assay, Expressing, Western Blot

Poxin transgene enhances the cytotoxic activity of NK-92 cells. A. Cytotoxicity of NK-92 cells with and without Poxin expression against K562 cells was measured over 4 h. B-C. Cells were co-cultured with target K562 cells for indicated times. B. Apoptosis levels in effector cells were assessed by FACS. C. Protein expression levels were detected by western blotting. (Representative data shown). D-G. Cells were harvested after 24 h in culture, and extracted total RNA was analyzed by RNA sequencing. (C = mock cells; P = Pox-NK-92 cells; n = 3). D. Heatmap. E. GO analysis. F. Differentially expressed genes in NK-92 and Pox-NK-92 groups were categorized by function. G. Top 10 upstream transcription factors and target genes were analyzed by network analysis. H. NK cell activation pathway was analyzed by western blotting. (Representative data shown). I. Quantitative analysis of Perforin expression level. J. mRNA levels of cytotoxic factors were analyzed by RT-qPCR (n = 3).

Journal: Journal of Advanced Research

Article Title: Enhancement of NK cell activity via DNA-sensing inhibition by Poxin transgene

doi: 10.1016/j.jare.2025.05.058

Figure Lengend Snippet: Poxin transgene enhances the cytotoxic activity of NK-92 cells. A. Cytotoxicity of NK-92 cells with and without Poxin expression against K562 cells was measured over 4 h. B-C. Cells were co-cultured with target K562 cells for indicated times. B. Apoptosis levels in effector cells were assessed by FACS. C. Protein expression levels were detected by western blotting. (Representative data shown). D-G. Cells were harvested after 24 h in culture, and extracted total RNA was analyzed by RNA sequencing. (C = mock cells; P = Pox-NK-92 cells; n = 3). D. Heatmap. E. GO analysis. F. Differentially expressed genes in NK-92 and Pox-NK-92 groups were categorized by function. G. Top 10 upstream transcription factors and target genes were analyzed by network analysis. H. NK cell activation pathway was analyzed by western blotting. (Representative data shown). I. Quantitative analysis of Perforin expression level. J. mRNA levels of cytotoxic factors were analyzed by RT-qPCR (n = 3).

Techniques: Activity Assay, Expressing, Cell Culture, Western Blot, RNA Sequencing, Activation Assay, Quantitative RT-PCR

Poxin enhances the anti-tumor capacity of NK-92 cells. A. Experimental scheme for mouse study (n = 4). B. Ventral bioluminescence (BLI) images following injection of NK92 or Pox-NK-92 cells. C. Quantification and statistical analysis (n = 4). D. Survival curves post-injection of K562-Luc cells. Data are presented as mean ± SEM. Mouse experiments were conducted independently twice, yielding consistent results.

Journal: Journal of Advanced Research

Article Title: Enhancement of NK cell activity via DNA-sensing inhibition by Poxin transgene

doi: 10.1016/j.jare.2025.05.058

Figure Lengend Snippet: Poxin enhances the anti-tumor capacity of NK-92 cells. A. Experimental scheme for mouse study (n = 4). B. Ventral bioluminescence (BLI) images following injection of NK92 or Pox-NK-92 cells. C. Quantification and statistical analysis (n = 4). D. Survival curves post-injection of K562-Luc cells. Data are presented as mean ± SEM. Mouse experiments were conducted independently twice, yielding consistent results.

Techniques: Injection

Poxin improves the anti-tumor efficacy of PD-L1-CAR-NK cells against H460 lung cancer. A. PD-L1-CAR expression levels in NK-92 cells. Abbreviations: TM = Transmembrane; Co-stim=Co-stimulatory; SD=Signaling domain. B. Cytotoxicity of CAR-NK-92 cells with or without Poxin expression against K562 and H460 cells, measured over 2 h. C. Experimental scheme for the H460 xenograft mouse model (n = 8). D. Tumor sizes were measured using digital calipers at the indicated time points. E. Representative tumor images from both groups at 28 days post-H460 inoculation. F. Tumor volumes were calculated. Error bars in panels B and D represent mean ± SEM. Mouse experiments were independently repeated twice with similar outcomes. Statistical significance was assessed using Two-way ANOVA (D) or Student's t -test (F).

Journal: Journal of Advanced Research

Article Title: Enhancement of NK cell activity via DNA-sensing inhibition by Poxin transgene

doi: 10.1016/j.jare.2025.05.058

Figure Lengend Snippet: Poxin improves the anti-tumor efficacy of PD-L1-CAR-NK cells against H460 lung cancer. A. PD-L1-CAR expression levels in NK-92 cells. Abbreviations: TM = Transmembrane; Co-stim=Co-stimulatory; SD=Signaling domain. B. Cytotoxicity of CAR-NK-92 cells with or without Poxin expression against K562 and H460 cells, measured over 2 h. C. Experimental scheme for the H460 xenograft mouse model (n = 8). D. Tumor sizes were measured using digital calipers at the indicated time points. E. Representative tumor images from both groups at 28 days post-H460 inoculation. F. Tumor volumes were calculated. Error bars in panels B and D represent mean ± SEM. Mouse experiments were independently repeated twice with similar outcomes. Statistical significance was assessed using Two-way ANOVA (D) or Student's t -test (F).

Techniques: Expressing

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1) Product Images from "Quantifying the Dual Effect of Antitumor and Pro-Tumor Human Neutrophils on Natural Killer Cell Behaviors in a Microphysiological System"

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