human pancreatic duct epithelial hpde cell line h6c7  (Absolute Biotech Inc)

Journal: Heliyon

Article Title: Sendai virus is robust and consistent in delivering genes into human pancreatic cancer cells

doi: 10.1016/j.heliyon.2024.e27221

Figure Lengend Snippet: The heterogeneous susceptibility to LV transduction among human pancreatic cancer epithelial cells. (A) The schematic diagram for the transduction of primary human PDAC with LV-GFP. (B–C) GFP expression in epithelial cells derived from freshly isolated human PDAC cells. MUC1 is a marker for PDAC. While most larger epithelial cells express GFP (B), smaller ductal epithelial cells barely express GFP (C).

Article Snippet: Human Pancreatic Duct Epithelial (HPDE) cell line H6c7 (Kerafast, Boston, MA # ECA001-FP) was maintained in Keratinocyte SFM medium (Thermo Fisher Scientific, Waltham, MA.

Techniques: Transduction, Expressing, Derivative Assay, Isolation, Marker

Journal: Heliyon

Article Title: Sendai virus is robust and consistent in delivering genes into human pancreatic cancer cells

doi: 10.1016/j.heliyon.2024.e27221

Figure Lengend Snippet: Transduction of human PDAC cells with LV-GFP. (A–B) Effect of LV-GFP transduction on the cell viability of PDAC cell lines (MIA PaCa-2, BxPC3, Panc-1, SW1990 (basal-like subtypes, circles) and CFPAC-1, Capan-1, and HPAF-II (classical subtypes, squares)) (A) and PDX-derived primary PDAC cells (ST-7599, ST-7270, and ST-14490) (B). Human normal fibroblasts (BJ6) and HPDE cells (H6c7) were used as controls. The percentage of live cells was calculated using D Horizon™ Fixable Viability Stain 660 by flow cytometry. Statistical significance was computed through linear regression model coefficients (p > 0.05, ). The asterisks in the figure refer to the P -value of linear regression model coefficients. * - P ≤ 0.05; ** - P ≤ 0.01; *** - P ≤ 0.001; **** - P ≤ 0.0001. ( C –D) Transduction efficiency of LV-GFP in PDAC cell lines (MIA PaCa-2, BxPC3, Panc-1, SW1990 (basal-like subtypes, circles) and CFPAC-1, Capan-1, and HPAF-II (classical subtypes, squares)) (C) and PDX-derived primary PDAC cells (ST-7599, ST-7270, and ST-14490) (D). BJ6 and H6c7 cells were used as controls. Functional infectious units per cell (IFU/cell) were calculated based on LV GFP titer in human fibrosarcoma HT1080 cells . The dependence of the fraction of GFP-expressing cells to the number of viral IFU/cell were fitted with a second-degree polynomial regression model. Statistical significance was computed through ANOVA to test whether there were any significant interactions of categorical variables (cell types) in the regression models estimating the relationships of the percentage of GFP-expressing cells to the number of viral IFU/cell used ( for p-value). The asterisks in the figure refer to the P -value of the ANOVA test. * - P ≤ 0.05; ** - P ≤ 0.01; *** - P ≤ 0.001; **** - P ≤ 0.0001. Each experiment was repeated at least three times independently (n ≥ 3). All data are represented as mean ± SD unless specified. SD bars can be smaller than the size of the symbols.

Article Snippet: Human Pancreatic Duct Epithelial (HPDE) cell line H6c7 (Kerafast, Boston, MA # ECA001-FP) was maintained in Keratinocyte SFM medium (Thermo Fisher Scientific, Waltham, MA.

Techniques: Transduction, Derivative Assay, Staining, Flow Cytometry, Functional Assay, Expressing

Journal: Heliyon

Article Title: Sendai virus is robust and consistent in delivering genes into human pancreatic cancer cells

doi: 10.1016/j.heliyon.2024.e27221

Figure Lengend Snippet: Transduction of human PDAC cells with SeV-GFP. (A–B) Effect on SeV-GFP transduction on the cell viability of PDAC cell lines (MIA PaCa-2, BxPC3, Panc-1, CFPAC-1, SW1990, Capan-1, and HPAF-II) (A) and PDX-derived primary PDAC cells (ST-7599, ST-7270, and ST-14490) (B). BJ6 fibroblast and H6c7 HPDE cells were used as normal controls. The percentage of live cells was calculated using D Horizon™ Fixable Viability Stain 660 by flow cytometry. Statistical significance was computed through linear regression model coefficients ( for p-values). ( C –D) Transduction efficiency of SeV-GFP in PDAC cell lines (MIA Paca-2, BxPC3, Panc-1, CFPAC-1, SW1990, Capan-1, and HPAF-II) (C) and PDX-derived primary PDAC cells (ST-7599, ST-7270, and ST-14490) (D). As controls, BJ6 and H6c7 cells were used. Functional infectious units per cell (IFU/cell) were calculated based on SeV-GFP titer in LLC-MK2 cells . The dependence of the fraction of GFP-expressing cells to the number of viral IFU/cell was fitted with a second-degree polynomial regression model. Statistical significance was computed through ANOVA to test whether there are any significant interactions of categorical variables (cell types) in the regression models estimating the relationships of the percentage of GFP-expressing cells to the number of viral IFU/cell used ( for p-value). The asterisks in the figure refer to the P -value of the ANOVA test. * - P ≤ 0.05; ** - P ≤ 0.01; *** - P ≤ 0.001; **** - P ≤ 0.0001. Each experiment was repeated at least three times independently (n ≥ 3). All data are represented as mean ± SD unless specified. SD bar can be smaller than the symbol's size.

Article Snippet: Human Pancreatic Duct Epithelial (HPDE) cell line H6c7 (Kerafast, Boston, MA # ECA001-FP) was maintained in Keratinocyte SFM medium (Thermo Fisher Scientific, Waltham, MA.

Techniques: Transduction, Derivative Assay, Staining, Flow Cytometry, Functional Assay, Expressing

Journal: Heliyon

Article Title: Sendai virus is robust and consistent in delivering genes into human pancreatic cancer cells

doi: 10.1016/j.heliyon.2024.e27221

Figure Lengend Snippet: Comparison of transduction efficiencies between LV-GFP and SeV-GFP. Transduction efficiencies are compared in control cell lines (HT1080, BJ6, and H6c7) (A), classical subtype PDAC lines (CFPAC-1, Capan-1, and HPAF-II) (B), basal-like subtype PDAC lines (MIA Paca-2, BxPC3, Panc-1, and SW1990) (C), and PDX-derived primary PDAC cells (ST-7599, ST-7270, and ST-14490) (D). The IFU/cell is calculated based on LV-GFP and SeV-GFP titer in corresponding cells. Each experiment was repeated at least three times independently (n ≥ 3). A second-degree polynomial regression model fitted the dependence of the fraction of GFP-expressing cells on the number of viral IFU/cell used. Statistical significance was computed through ANOVA to test whether there are any significant interactions of categorical variables (viral vector type) in the regression models estimating the relationships of the percentage of GFP expressing cells to the number of viral IFU/cell used ( for p-value). The asterisks in the figure refer to the P -value of the ANOVA test. * - P ≤ 0.05; ** - P ≤ 0.01; *** - P ≤ 0.001; **** - P ≤ 0.0001. All data are represented as mean ± SD unless specified. An SD bar can be smaller than the symbol's size.

Article Snippet: Human Pancreatic Duct Epithelial (HPDE) cell line H6c7 (Kerafast, Boston, MA # ECA001-FP) was maintained in Keratinocyte SFM medium (Thermo Fisher Scientific, Waltham, MA.

Techniques: Comparison, Transduction, Derivative Assay, Expressing, Plasmid Preparation

Journal: Heliyon

Article Title: Sendai virus is robust and consistent in delivering genes into human pancreatic cancer cells

doi: 10.1016/j.heliyon.2024.e27221

Figure Lengend Snippet: Relative transduction efficiencies of LV-GFP and SeV-GFP in PDAC cell lines and PDX-derived primary PDAC cells. The relative transduction efficiencies of LV and SeV vectors in PDAC cells (cell lines or primary cells) were calculated by normalizing titers obtained from PDAC cells to the titers obtained on the control H6c7 HPDE cells (n = 3). Statistical significance was computed with the nonparametric Mann-Whitney U test. The relative transduction efficiency of SeV-GFP was significantly higher than that of LV-GFP across all tested PDAC cells (Mann Whitney U test, p < 0.05 ). The relative transduction efficiency of LV-GFP was significantly lower in the classical subtype than in the basal-like subtype of PDAC (Mann Whitney U test, p = 0.04 ). In contrast, there was no difference in the relative transduction efficiencies of SeV-GFP between classical and basal-like subtypes of PDAC cells (Mann Whitney U test, p = 1 ). Basal-like subtype PDAC lines (MIA Paca-2, BxPC3, Panc-1, and SW1990); classical subtype PDAC lines (CFPAC-1; Capan-1; HPAF-II; Primary PDAC cell (ST-7599; ST-7270; ST-14490); foreskin fibroblast BJ6; HPDE cell (H6c7). Since the presented data are not the result of direct measurements and do not satisfy all propagation of error requirements , the standard deviations are not shown.

Article Snippet: Human Pancreatic Duct Epithelial (HPDE) cell line H6c7 (Kerafast, Boston, MA # ECA001-FP) was maintained in Keratinocyte SFM medium (Thermo Fisher Scientific, Waltham, MA.

Techniques: Transduction, Derivative Assay, MANN-WHITNEY