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envelope hepg2 env gene  (ATCC)


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    ATCC envelope hepg2 env gene
    (A) Timeline of notable events during a representative 11-hour live-imaging assay (~7-minute acquisition intervals; experiment performed twice), in which engineered HBV Env183-191–specific T cells (TCRe–T cells) were introduced into the device containing GFP-expressing <t>HepG2-Env</t> cells cultured in a 3D collagen matrix. Engineered T cells were labeled with CellTracker BMQC (blue), while DRAQ7 (red) was added in the culture media. HepG2-Env target cells are shown in green (GFP). The magnified maximum intensity projections of a single HepG2-Env cell are shown at the indicated times. Scale bar: 10 μM. (B) Representative maximum intensity projections of a region of the collagen gel showing HepG2-Env cells at 0 and 15 hours after incubation alone, with the addition of 10% DMSO, or with engineered TCRe–T cells. The mean fluorescence intensity (MFI) of GFP and DRAQ7 of each HepG2-Env target cell identified in Imaris was plotted at 0 and 15 hours after incubation at the respective conditions. Devices in which HepG2-Env cells were cultured with DMSO (red) were plotted in the background for reference, with the percentage of dead target cells quantified in devices with (blue) or without (green) the addition of TCRe–T cells at time points shown. The graph shows the percentage of killing quantified for the respective conditions; each point represents an individual experiment. Original magnification, ×10. (C) Representative maximum intensity projections of a region of the collagen gel showing HepG2-Env cells at 0 and 15 hours after incubation with engineered TCRe–T cells or Core18-27–specific T cells (TCRc–T cells). The mean percentage of killed target cells after overnight incubation without T cells or with engineered TCRe- or TCRc- specific T cells at the antigen-specific E/T ratios are shown as bars; each point represents an individual experiment. All experiments were repeated in at least 3 microfluidic devices. The amount of target cells killed when TCRe–T cells were analyzed using a 2D well-based killing assay is shown for comparison. Original magnification, ×10. (D) 41 secreted factors were quantified using multiplex bead-based assay. Factors with detected concentrations less than 10 pg/ml in all samples were removed from analysis. The mean concentration of secreted factors present in the supernatant retrieved from microdevices in which only HepG2-Env target cells were seeded (n = 4) or cocultured with retroviral-transduced TCRe–T cells (n = 2).
    Envelope Hepg2 Env Gene, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 31326 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/envelope hepg2 env gene/product/ATCC
    Average 99 stars, based on 31326 article reviews
    envelope hepg2 env gene - by Bioz Stars, 2026-02
    99/100 stars

    Images

    1) Product Images from "A 3D microfluidic model for preclinical evaluation of TCR-engineered T cells against solid tumors"

    Article Title: A 3D microfluidic model for preclinical evaluation of TCR-engineered T cells against solid tumors

    Journal: JCI Insight

    doi: 10.1172/jci.insight.89762

    (A) Timeline of notable events during a representative 11-hour live-imaging assay (~7-minute acquisition intervals; experiment performed twice), in which engineered HBV Env183-191–specific T cells (TCRe–T cells) were introduced into the device containing GFP-expressing HepG2-Env cells cultured in a 3D collagen matrix. Engineered T cells were labeled with CellTracker BMQC (blue), while DRAQ7 (red) was added in the culture media. HepG2-Env target cells are shown in green (GFP). The magnified maximum intensity projections of a single HepG2-Env cell are shown at the indicated times. Scale bar: 10 μM. (B) Representative maximum intensity projections of a region of the collagen gel showing HepG2-Env cells at 0 and 15 hours after incubation alone, with the addition of 10% DMSO, or with engineered TCRe–T cells. The mean fluorescence intensity (MFI) of GFP and DRAQ7 of each HepG2-Env target cell identified in Imaris was plotted at 0 and 15 hours after incubation at the respective conditions. Devices in which HepG2-Env cells were cultured with DMSO (red) were plotted in the background for reference, with the percentage of dead target cells quantified in devices with (blue) or without (green) the addition of TCRe–T cells at time points shown. The graph shows the percentage of killing quantified for the respective conditions; each point represents an individual experiment. Original magnification, ×10. (C) Representative maximum intensity projections of a region of the collagen gel showing HepG2-Env cells at 0 and 15 hours after incubation with engineered TCRe–T cells or Core18-27–specific T cells (TCRc–T cells). The mean percentage of killed target cells after overnight incubation without T cells or with engineered TCRe- or TCRc- specific T cells at the antigen-specific E/T ratios are shown as bars; each point represents an individual experiment. All experiments were repeated in at least 3 microfluidic devices. The amount of target cells killed when TCRe–T cells were analyzed using a 2D well-based killing assay is shown for comparison. Original magnification, ×10. (D) 41 secreted factors were quantified using multiplex bead-based assay. Factors with detected concentrations less than 10 pg/ml in all samples were removed from analysis. The mean concentration of secreted factors present in the supernatant retrieved from microdevices in which only HepG2-Env target cells were seeded (n = 4) or cocultured with retroviral-transduced TCRe–T cells (n = 2).
    Figure Legend Snippet: (A) Timeline of notable events during a representative 11-hour live-imaging assay (~7-minute acquisition intervals; experiment performed twice), in which engineered HBV Env183-191–specific T cells (TCRe–T cells) were introduced into the device containing GFP-expressing HepG2-Env cells cultured in a 3D collagen matrix. Engineered T cells were labeled with CellTracker BMQC (blue), while DRAQ7 (red) was added in the culture media. HepG2-Env target cells are shown in green (GFP). The magnified maximum intensity projections of a single HepG2-Env cell are shown at the indicated times. Scale bar: 10 μM. (B) Representative maximum intensity projections of a region of the collagen gel showing HepG2-Env cells at 0 and 15 hours after incubation alone, with the addition of 10% DMSO, or with engineered TCRe–T cells. The mean fluorescence intensity (MFI) of GFP and DRAQ7 of each HepG2-Env target cell identified in Imaris was plotted at 0 and 15 hours after incubation at the respective conditions. Devices in which HepG2-Env cells were cultured with DMSO (red) were plotted in the background for reference, with the percentage of dead target cells quantified in devices with (blue) or without (green) the addition of TCRe–T cells at time points shown. The graph shows the percentage of killing quantified for the respective conditions; each point represents an individual experiment. Original magnification, ×10. (C) Representative maximum intensity projections of a region of the collagen gel showing HepG2-Env cells at 0 and 15 hours after incubation with engineered TCRe–T cells or Core18-27–specific T cells (TCRc–T cells). The mean percentage of killed target cells after overnight incubation without T cells or with engineered TCRe- or TCRc- specific T cells at the antigen-specific E/T ratios are shown as bars; each point represents an individual experiment. All experiments were repeated in at least 3 microfluidic devices. The amount of target cells killed when TCRe–T cells were analyzed using a 2D well-based killing assay is shown for comparison. Original magnification, ×10. (D) 41 secreted factors were quantified using multiplex bead-based assay. Factors with detected concentrations less than 10 pg/ml in all samples were removed from analysis. The mean concentration of secreted factors present in the supernatant retrieved from microdevices in which only HepG2-Env target cells were seeded (n = 4) or cocultured with retroviral-transduced TCRe–T cells (n = 2).

    Techniques Used: Imaging, Expressing, Cell Culture, Labeling, Incubation, Fluorescence, Multiplex Assay, Bead-based Assay, Concentration Assay

    (A) Schematic illustrating the 3D volume reconstruction of HepG2-Env aggregates embedded in the collagen gel of the microdevice (scale bar: 100 μm [left]; 50 μm [middle and right]). (B) Timeline depicting a representative 15.5-hour live-imaging assay (~30-minute acquisition intervals; experiment performed twice), in which engineered HBV Env183-191–specific T cells were introduced into the device containing GFP-expressing HepG2-Env aggregates cultured in a 3D collagen matrix. DRAQ7 was added in the culture media labeling dead cells red. The reconstructed aggregates were shown at the indicated times (scale bar: 50 μm). (C) The density of engineered T cells randomly moving in empty collagen gel regions (noninteracting) and within HepG2-Env aggregates (interacting) was quantified in 2 live-imaging experiments depicted. (D) Representative reconstructed aggregates at 0 hours and after overnight incubation alone (left), with the addition of 10% DMSO or with engineered HBV-specific T cells (scale bar: 50 μm). The summary of the percentage of dead aggregates was quantified for the respective conditions. Each dot represents a single experiment. Scale bar: 50 μM. Statistical significance was evaluated with 2-tailed t test.
    Figure Legend Snippet: (A) Schematic illustrating the 3D volume reconstruction of HepG2-Env aggregates embedded in the collagen gel of the microdevice (scale bar: 100 μm [left]; 50 μm [middle and right]). (B) Timeline depicting a representative 15.5-hour live-imaging assay (~30-minute acquisition intervals; experiment performed twice), in which engineered HBV Env183-191–specific T cells were introduced into the device containing GFP-expressing HepG2-Env aggregates cultured in a 3D collagen matrix. DRAQ7 was added in the culture media labeling dead cells red. The reconstructed aggregates were shown at the indicated times (scale bar: 50 μm). (C) The density of engineered T cells randomly moving in empty collagen gel regions (noninteracting) and within HepG2-Env aggregates (interacting) was quantified in 2 live-imaging experiments depicted. (D) Representative reconstructed aggregates at 0 hours and after overnight incubation alone (left), with the addition of 10% DMSO or with engineered HBV-specific T cells (scale bar: 50 μm). The summary of the percentage of dead aggregates was quantified for the respective conditions. Each dot represents a single experiment. Scale bar: 50 μM. Statistical significance was evaluated with 2-tailed t test.

    Techniques Used: Imaging, Expressing, Cell Culture, Labeling, Incubation

    (A) Spontaneous death of HepG2-Env target cells cultured under specified conditions. (B) Normalized percentage of killed target cells when retroviral-transduced (left) or activated mRNA-electroporated (right) TCR-engineered T cells were cocultured with dispersed HepG2-Env target cells under specified conditions. Results were normalized to data obtained under typical in vitro assay conditions (20% oxygen and no inflammatory cytokines). Each point represents a single experiment. Outliers are indicated in orange. Statistical significance was evaluated with 2-way ANOVA with Tukey’s multiple comparison test. (C) The percentage of dead HepG2-Env aggregates before and after overnight coculture with retroviral-transduced (left), activated nonelectroporated (middle), or mRNA-electroporated (right) TCR-engineered T cells under specified conditions. Each dot represents a single experiment. Statistical significance was evaluated with 2-tailed t test.
    Figure Legend Snippet: (A) Spontaneous death of HepG2-Env target cells cultured under specified conditions. (B) Normalized percentage of killed target cells when retroviral-transduced (left) or activated mRNA-electroporated (right) TCR-engineered T cells were cocultured with dispersed HepG2-Env target cells under specified conditions. Results were normalized to data obtained under typical in vitro assay conditions (20% oxygen and no inflammatory cytokines). Each point represents a single experiment. Outliers are indicated in orange. Statistical significance was evaluated with 2-way ANOVA with Tukey’s multiple comparison test. (C) The percentage of dead HepG2-Env aggregates before and after overnight coculture with retroviral-transduced (left), activated nonelectroporated (middle), or mRNA-electroporated (right) TCR-engineered T cells under specified conditions. Each dot represents a single experiment. Statistical significance was evaluated with 2-tailed t test.

    Techniques Used: Cell Culture, In Vitro

    (A) Normalized percentage of killed target cells when retroV-TCRe (left) or mRNA-TCRe (right) T cells were cocultured with luciferase+ HepG2-Env target cells under specified conditions. Results were normalized to data obtained under typical in vitro assay conditions. Data shown were compiled from experiments with 2 different E/T ratios performed in duplicates, as denoted in Supplemental Figure 2. Statistical significance was evaluated with 2-way ANOVA with Tukey’s multiple comparison test. (B) Representative images of 3D microdevices in which mRNA TCRe–T cells were cultured overnight with HepG2-Env target cells in the respective conditions. Green spheres denote the living HepG2-Env target cells, while red spheres denotes the mRNA TCRe–T cells. The number of T cells present in the gel at the end of the experiment (each dot represents the number of T cells in each microdevice) and the position of all gel-invading T cells in each condition (each dot represent the position of a T cell) are shown. Statistical significance was evaluated with 2-tailed t test.
    Figure Legend Snippet: (A) Normalized percentage of killed target cells when retroV-TCRe (left) or mRNA-TCRe (right) T cells were cocultured with luciferase+ HepG2-Env target cells under specified conditions. Results were normalized to data obtained under typical in vitro assay conditions. Data shown were compiled from experiments with 2 different E/T ratios performed in duplicates, as denoted in Supplemental Figure 2. Statistical significance was evaluated with 2-way ANOVA with Tukey’s multiple comparison test. (B) Representative images of 3D microdevices in which mRNA TCRe–T cells were cultured overnight with HepG2-Env target cells in the respective conditions. Green spheres denote the living HepG2-Env target cells, while red spheres denotes the mRNA TCRe–T cells. The number of T cells present in the gel at the end of the experiment (each dot represents the number of T cells in each microdevice) and the position of all gel-invading T cells in each condition (each dot represent the position of a T cell) are shown. Statistical significance was evaluated with 2-tailed t test.

    Techniques Used: Luciferase, In Vitro, Cell Culture

    (A) Cytotoxicity of retroV-TCR–T cells was analyzed in the presence of increasing concentrations of immunosuppressive mTOR inhibitor (rapamycin). HepG2-Env target cells were seeded as before, and rapamycin was introduced with the TCR–T cells. Each dot represents a single experiment. Statistical significance was evaluated with 2-way ANOVA with Tukey’s multiple comparison test. (B) Cytotoxicity of retroV-TCR–T cells against HepG2-Env target cells pretreated for 2 weeks with 5 nM rapamycin was analyzed. Untreated targets were used as controls. Rapamycin concentrations were maintained throughout the experiment to mimic the in vivo condition of a patient under stable immunosuppression. Each dot represents a single experiment. Statistical significance was evaluated with 2-tailed t test.
    Figure Legend Snippet: (A) Cytotoxicity of retroV-TCR–T cells was analyzed in the presence of increasing concentrations of immunosuppressive mTOR inhibitor (rapamycin). HepG2-Env target cells were seeded as before, and rapamycin was introduced with the TCR–T cells. Each dot represents a single experiment. Statistical significance was evaluated with 2-way ANOVA with Tukey’s multiple comparison test. (B) Cytotoxicity of retroV-TCR–T cells against HepG2-Env target cells pretreated for 2 weeks with 5 nM rapamycin was analyzed. Untreated targets were used as controls. Rapamycin concentrations were maintained throughout the experiment to mimic the in vivo condition of a patient under stable immunosuppression. Each dot represents a single experiment. Statistical significance was evaluated with 2-tailed t test.

    Techniques Used: In Vivo



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    envelope hepg2 env gene  (ATCC)
    99
    ATCC envelope hepg2 env gene
    (A) Timeline of notable events during a representative 11-hour live-imaging assay (~7-minute acquisition intervals; experiment performed twice), in which engineered HBV Env183-191–specific T cells (TCRe–T cells) were introduced into the device containing GFP-expressing <t>HepG2-Env</t> cells cultured in a 3D collagen matrix. Engineered T cells were labeled with CellTracker BMQC (blue), while DRAQ7 (red) was added in the culture media. HepG2-Env target cells are shown in green (GFP). The magnified maximum intensity projections of a single HepG2-Env cell are shown at the indicated times. Scale bar: 10 μM. (B) Representative maximum intensity projections of a region of the collagen gel showing HepG2-Env cells at 0 and 15 hours after incubation alone, with the addition of 10% DMSO, or with engineered TCRe–T cells. The mean fluorescence intensity (MFI) of GFP and DRAQ7 of each HepG2-Env target cell identified in Imaris was plotted at 0 and 15 hours after incubation at the respective conditions. Devices in which HepG2-Env cells were cultured with DMSO (red) were plotted in the background for reference, with the percentage of dead target cells quantified in devices with (blue) or without (green) the addition of TCRe–T cells at time points shown. The graph shows the percentage of killing quantified for the respective conditions; each point represents an individual experiment. Original magnification, ×10. (C) Representative maximum intensity projections of a region of the collagen gel showing HepG2-Env cells at 0 and 15 hours after incubation with engineered TCRe–T cells or Core18-27–specific T cells (TCRc–T cells). The mean percentage of killed target cells after overnight incubation without T cells or with engineered TCRe- or TCRc- specific T cells at the antigen-specific E/T ratios are shown as bars; each point represents an individual experiment. All experiments were repeated in at least 3 microfluidic devices. The amount of target cells killed when TCRe–T cells were analyzed using a 2D well-based killing assay is shown for comparison. Original magnification, ×10. (D) 41 secreted factors were quantified using multiplex bead-based assay. Factors with detected concentrations less than 10 pg/ml in all samples were removed from analysis. The mean concentration of secreted factors present in the supernatant retrieved from microdevices in which only HepG2-Env target cells were seeded (n = 4) or cocultured with retroviral-transduced TCRe–T cells (n = 2).
    Envelope Hepg2 Env Gene, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/envelope hepg2 env gene/product/ATCC
    Average 99 stars, based on 1 article reviews
    envelope hepg2 env gene - by Bioz Stars, 2026-02
    99/100 stars
    		  Buy from Supplier

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    (A) Timeline of notable events during a representative 11-hour live-imaging assay (~7-minute acquisition intervals; experiment performed twice), in which engineered HBV Env183-191–specific T cells (TCRe–T cells) were introduced into the device containing GFP-expressing HepG2-Env cells cultured in a 3D collagen matrix. Engineered T cells were labeled with CellTracker BMQC (blue), while DRAQ7 (red) was added in the culture media. HepG2-Env target cells are shown in green (GFP). The magnified maximum intensity projections of a single HepG2-Env cell are shown at the indicated times. Scale bar: 10 μM. (B) Representative maximum intensity projections of a region of the collagen gel showing HepG2-Env cells at 0 and 15 hours after incubation alone, with the addition of 10% DMSO, or with engineered TCRe–T cells. The mean fluorescence intensity (MFI) of GFP and DRAQ7 of each HepG2-Env target cell identified in Imaris was plotted at 0 and 15 hours after incubation at the respective conditions. Devices in which HepG2-Env cells were cultured with DMSO (red) were plotted in the background for reference, with the percentage of dead target cells quantified in devices with (blue) or without (green) the addition of TCRe–T cells at time points shown. The graph shows the percentage of killing quantified for the respective conditions; each point represents an individual experiment. Original magnification, ×10. (C) Representative maximum intensity projections of a region of the collagen gel showing HepG2-Env cells at 0 and 15 hours after incubation with engineered TCRe–T cells or Core18-27–specific T cells (TCRc–T cells). The mean percentage of killed target cells after overnight incubation without T cells or with engineered TCRe- or TCRc- specific T cells at the antigen-specific E/T ratios are shown as bars; each point represents an individual experiment. All experiments were repeated in at least 3 microfluidic devices. The amount of target cells killed when TCRe–T cells were analyzed using a 2D well-based killing assay is shown for comparison. Original magnification, ×10. (D) 41 secreted factors were quantified using multiplex bead-based assay. Factors with detected concentrations less than 10 pg/ml in all samples were removed from analysis. The mean concentration of secreted factors present in the supernatant retrieved from microdevices in which only HepG2-Env target cells were seeded (n = 4) or cocultured with retroviral-transduced TCRe–T cells (n = 2).

    Journal: JCI Insight

    Article Title: A 3D microfluidic model for preclinical evaluation of TCR-engineered T cells against solid tumors

    doi: 10.1172/jci.insight.89762

    Figure Lengend Snippet: (A) Timeline of notable events during a representative 11-hour live-imaging assay (~7-minute acquisition intervals; experiment performed twice), in which engineered HBV Env183-191–specific T cells (TCRe–T cells) were introduced into the device containing GFP-expressing HepG2-Env cells cultured in a 3D collagen matrix. Engineered T cells were labeled with CellTracker BMQC (blue), while DRAQ7 (red) was added in the culture media. HepG2-Env target cells are shown in green (GFP). The magnified maximum intensity projections of a single HepG2-Env cell are shown at the indicated times. Scale bar: 10 μM. (B) Representative maximum intensity projections of a region of the collagen gel showing HepG2-Env cells at 0 and 15 hours after incubation alone, with the addition of 10% DMSO, or with engineered TCRe–T cells. The mean fluorescence intensity (MFI) of GFP and DRAQ7 of each HepG2-Env target cell identified in Imaris was plotted at 0 and 15 hours after incubation at the respective conditions. Devices in which HepG2-Env cells were cultured with DMSO (red) were plotted in the background for reference, with the percentage of dead target cells quantified in devices with (blue) or without (green) the addition of TCRe–T cells at time points shown. The graph shows the percentage of killing quantified for the respective conditions; each point represents an individual experiment. Original magnification, ×10. (C) Representative maximum intensity projections of a region of the collagen gel showing HepG2-Env cells at 0 and 15 hours after incubation with engineered TCRe–T cells or Core18-27–specific T cells (TCRc–T cells). The mean percentage of killed target cells after overnight incubation without T cells or with engineered TCRe- or TCRc- specific T cells at the antigen-specific E/T ratios are shown as bars; each point represents an individual experiment. All experiments were repeated in at least 3 microfluidic devices. The amount of target cells killed when TCRe–T cells were analyzed using a 2D well-based killing assay is shown for comparison. Original magnification, ×10. (D) 41 secreted factors were quantified using multiplex bead-based assay. Factors with detected concentrations less than 10 pg/ml in all samples were removed from analysis. The mean concentration of secreted factors present in the supernatant retrieved from microdevices in which only HepG2-Env target cells were seeded (n = 4) or cocultured with retroviral-transduced TCRe–T cells (n = 2).

    Article Snippet: HBV antigen–expressing HepG2 target cells The human liver carcinoma cell line, HepG2 (ATCC), was transduced with a construct containing either the full genotype D HBV core (HepG2-Core) or envelope (HepG2-Env) gene covalently linked to GFP using the Lenti-X HTX packaging system (Clontech) according to the manufacturer’s instructions.

    Techniques: Imaging, Expressing, Cell Culture, Labeling, Incubation, Fluorescence, Multiplex Assay, Bead-based Assay, Concentration Assay

    (A) Schematic illustrating the 3D volume reconstruction of HepG2-Env aggregates embedded in the collagen gel of the microdevice (scale bar: 100 μm [left]; 50 μm [middle and right]). (B) Timeline depicting a representative 15.5-hour live-imaging assay (~30-minute acquisition intervals; experiment performed twice), in which engineered HBV Env183-191–specific T cells were introduced into the device containing GFP-expressing HepG2-Env aggregates cultured in a 3D collagen matrix. DRAQ7 was added in the culture media labeling dead cells red. The reconstructed aggregates were shown at the indicated times (scale bar: 50 μm). (C) The density of engineered T cells randomly moving in empty collagen gel regions (noninteracting) and within HepG2-Env aggregates (interacting) was quantified in 2 live-imaging experiments depicted. (D) Representative reconstructed aggregates at 0 hours and after overnight incubation alone (left), with the addition of 10% DMSO or with engineered HBV-specific T cells (scale bar: 50 μm). The summary of the percentage of dead aggregates was quantified for the respective conditions. Each dot represents a single experiment. Scale bar: 50 μM. Statistical significance was evaluated with 2-tailed t test.

    Journal: JCI Insight

    Article Title: A 3D microfluidic model for preclinical evaluation of TCR-engineered T cells against solid tumors

    doi: 10.1172/jci.insight.89762

    Figure Lengend Snippet: (A) Schematic illustrating the 3D volume reconstruction of HepG2-Env aggregates embedded in the collagen gel of the microdevice (scale bar: 100 μm [left]; 50 μm [middle and right]). (B) Timeline depicting a representative 15.5-hour live-imaging assay (~30-minute acquisition intervals; experiment performed twice), in which engineered HBV Env183-191–specific T cells were introduced into the device containing GFP-expressing HepG2-Env aggregates cultured in a 3D collagen matrix. DRAQ7 was added in the culture media labeling dead cells red. The reconstructed aggregates were shown at the indicated times (scale bar: 50 μm). (C) The density of engineered T cells randomly moving in empty collagen gel regions (noninteracting) and within HepG2-Env aggregates (interacting) was quantified in 2 live-imaging experiments depicted. (D) Representative reconstructed aggregates at 0 hours and after overnight incubation alone (left), with the addition of 10% DMSO or with engineered HBV-specific T cells (scale bar: 50 μm). The summary of the percentage of dead aggregates was quantified for the respective conditions. Each dot represents a single experiment. Scale bar: 50 μM. Statistical significance was evaluated with 2-tailed t test.

    Article Snippet: HBV antigen–expressing HepG2 target cells The human liver carcinoma cell line, HepG2 (ATCC), was transduced with a construct containing either the full genotype D HBV core (HepG2-Core) or envelope (HepG2-Env) gene covalently linked to GFP using the Lenti-X HTX packaging system (Clontech) according to the manufacturer’s instructions.

    Techniques: Imaging, Expressing, Cell Culture, Labeling, Incubation

    (A) Spontaneous death of HepG2-Env target cells cultured under specified conditions. (B) Normalized percentage of killed target cells when retroviral-transduced (left) or activated mRNA-electroporated (right) TCR-engineered T cells were cocultured with dispersed HepG2-Env target cells under specified conditions. Results were normalized to data obtained under typical in vitro assay conditions (20% oxygen and no inflammatory cytokines). Each point represents a single experiment. Outliers are indicated in orange. Statistical significance was evaluated with 2-way ANOVA with Tukey’s multiple comparison test. (C) The percentage of dead HepG2-Env aggregates before and after overnight coculture with retroviral-transduced (left), activated nonelectroporated (middle), or mRNA-electroporated (right) TCR-engineered T cells under specified conditions. Each dot represents a single experiment. Statistical significance was evaluated with 2-tailed t test.

    Journal: JCI Insight

    Article Title: A 3D microfluidic model for preclinical evaluation of TCR-engineered T cells against solid tumors

    doi: 10.1172/jci.insight.89762

    Figure Lengend Snippet: (A) Spontaneous death of HepG2-Env target cells cultured under specified conditions. (B) Normalized percentage of killed target cells when retroviral-transduced (left) or activated mRNA-electroporated (right) TCR-engineered T cells were cocultured with dispersed HepG2-Env target cells under specified conditions. Results were normalized to data obtained under typical in vitro assay conditions (20% oxygen and no inflammatory cytokines). Each point represents a single experiment. Outliers are indicated in orange. Statistical significance was evaluated with 2-way ANOVA with Tukey’s multiple comparison test. (C) The percentage of dead HepG2-Env aggregates before and after overnight coculture with retroviral-transduced (left), activated nonelectroporated (middle), or mRNA-electroporated (right) TCR-engineered T cells under specified conditions. Each dot represents a single experiment. Statistical significance was evaluated with 2-tailed t test.

    Article Snippet: HBV antigen–expressing HepG2 target cells The human liver carcinoma cell line, HepG2 (ATCC), was transduced with a construct containing either the full genotype D HBV core (HepG2-Core) or envelope (HepG2-Env) gene covalently linked to GFP using the Lenti-X HTX packaging system (Clontech) according to the manufacturer’s instructions.

    Techniques: Cell Culture, In Vitro

    (A) Normalized percentage of killed target cells when retroV-TCRe (left) or mRNA-TCRe (right) T cells were cocultured with luciferase+ HepG2-Env target cells under specified conditions. Results were normalized to data obtained under typical in vitro assay conditions. Data shown were compiled from experiments with 2 different E/T ratios performed in duplicates, as denoted in Supplemental Figure 2. Statistical significance was evaluated with 2-way ANOVA with Tukey’s multiple comparison test. (B) Representative images of 3D microdevices in which mRNA TCRe–T cells were cultured overnight with HepG2-Env target cells in the respective conditions. Green spheres denote the living HepG2-Env target cells, while red spheres denotes the mRNA TCRe–T cells. The number of T cells present in the gel at the end of the experiment (each dot represents the number of T cells in each microdevice) and the position of all gel-invading T cells in each condition (each dot represent the position of a T cell) are shown. Statistical significance was evaluated with 2-tailed t test.

    Journal: JCI Insight

    Article Title: A 3D microfluidic model for preclinical evaluation of TCR-engineered T cells against solid tumors

    doi: 10.1172/jci.insight.89762

    Figure Lengend Snippet: (A) Normalized percentage of killed target cells when retroV-TCRe (left) or mRNA-TCRe (right) T cells were cocultured with luciferase+ HepG2-Env target cells under specified conditions. Results were normalized to data obtained under typical in vitro assay conditions. Data shown were compiled from experiments with 2 different E/T ratios performed in duplicates, as denoted in Supplemental Figure 2. Statistical significance was evaluated with 2-way ANOVA with Tukey’s multiple comparison test. (B) Representative images of 3D microdevices in which mRNA TCRe–T cells were cultured overnight with HepG2-Env target cells in the respective conditions. Green spheres denote the living HepG2-Env target cells, while red spheres denotes the mRNA TCRe–T cells. The number of T cells present in the gel at the end of the experiment (each dot represents the number of T cells in each microdevice) and the position of all gel-invading T cells in each condition (each dot represent the position of a T cell) are shown. Statistical significance was evaluated with 2-tailed t test.

    Article Snippet: HBV antigen–expressing HepG2 target cells The human liver carcinoma cell line, HepG2 (ATCC), was transduced with a construct containing either the full genotype D HBV core (HepG2-Core) or envelope (HepG2-Env) gene covalently linked to GFP using the Lenti-X HTX packaging system (Clontech) according to the manufacturer’s instructions.

    Techniques: Luciferase, In Vitro, Cell Culture

    (A) Cytotoxicity of retroV-TCR–T cells was analyzed in the presence of increasing concentrations of immunosuppressive mTOR inhibitor (rapamycin). HepG2-Env target cells were seeded as before, and rapamycin was introduced with the TCR–T cells. Each dot represents a single experiment. Statistical significance was evaluated with 2-way ANOVA with Tukey’s multiple comparison test. (B) Cytotoxicity of retroV-TCR–T cells against HepG2-Env target cells pretreated for 2 weeks with 5 nM rapamycin was analyzed. Untreated targets were used as controls. Rapamycin concentrations were maintained throughout the experiment to mimic the in vivo condition of a patient under stable immunosuppression. Each dot represents a single experiment. Statistical significance was evaluated with 2-tailed t test.

    Journal: JCI Insight

    Article Title: A 3D microfluidic model for preclinical evaluation of TCR-engineered T cells against solid tumors

    doi: 10.1172/jci.insight.89762

    Figure Lengend Snippet: (A) Cytotoxicity of retroV-TCR–T cells was analyzed in the presence of increasing concentrations of immunosuppressive mTOR inhibitor (rapamycin). HepG2-Env target cells were seeded as before, and rapamycin was introduced with the TCR–T cells. Each dot represents a single experiment. Statistical significance was evaluated with 2-way ANOVA with Tukey’s multiple comparison test. (B) Cytotoxicity of retroV-TCR–T cells against HepG2-Env target cells pretreated for 2 weeks with 5 nM rapamycin was analyzed. Untreated targets were used as controls. Rapamycin concentrations were maintained throughout the experiment to mimic the in vivo condition of a patient under stable immunosuppression. Each dot represents a single experiment. Statistical significance was evaluated with 2-tailed t test.

    Article Snippet: HBV antigen–expressing HepG2 target cells The human liver carcinoma cell line, HepG2 (ATCC), was transduced with a construct containing either the full genotype D HBV core (HepG2-Core) or envelope (HepG2-Env) gene covalently linked to GFP using the Lenti-X HTX packaging system (Clontech) according to the manufacturer’s instructions.

    Techniques: In Vivo