Journal: iScience

Article Title: Third-generation taxanes SB-T-121605 and SB-T-121606 are effective in pancreatic ductal adenocarcinoma

doi: 10.1016/j.isci.2024.109044

Figure Lengend Snippet: In vitro efficacy of paclitaxel and SB-Ts (A) Graph showing the cell viability (% of control cells) of Paca-44 and BxPC-3 cell line treated with paclitaxel (PTX), second- and third-generation of SB-Ts for 72 h. Table with IC 50 values as mean ± S.D. of three independent experiments presented. (B) Representative graphs depicting the effect of concentrations corresponding to IC 50 (5–30 nM) 100 nM and 300 nM of PTX, SB-T-121605, and SB-T-12606 on cellular proliferation in Paca-44 and BxPC-3 cells. The data are presented as mean ± standard deviation of two independent experiments.

Article Snippet: BxPC3 , Cell Lines Service GmbH Company, Eppelheim, Baden- Dr.-Eckener-Str. 8, Eppelheim, Baden-Württemberg, 69214, Germany , Product number: 305031.

Techniques: In Vitro, Control, Standard Deviation

Journal: iScience

Article Title: Third-generation taxanes SB-T-121605 and SB-T-121606 are effective in pancreatic ductal adenocarcinoma

doi: 10.1016/j.isci.2024.109044

Figure Lengend Snippet: Cell cycle changes after 24 h of incubation of cells with paclitaxel and experimental SB-Ts in vitro (A and B) Taxane concentrations corresponding to IC 50 values (5–30 nM) and 100 nM concentration were used in (A) Paca-44, (B) BxPC-3 cell line. Histograms display three independent experiments as mean ± SD.

Article Snippet: BxPC3 , Cell Lines Service GmbH Company, Eppelheim, Baden- Dr.-Eckener-Str. 8, Eppelheim, Baden-Württemberg, 69214, Germany , Product number: 305031.

Techniques: Incubation, In Vitro, Concentration Assay

Journal: iScience

Article Title: Third-generation taxanes SB-T-121605 and SB-T-121606 are effective in pancreatic ductal adenocarcinoma

doi: 10.1016/j.isci.2024.109044

Figure Lengend Snippet: Western blot analysis of phospho-Bcl-2 (P-Bcl-2), PARP and cleaved caspase-3 expression in Paca-44 and BxPC-3 cells after paclitaxel and SB-Ts treatment (A) Cells treated with 25 nM (BxPC-3) or 30 nM (Paca-44) PTX, 5 nM SB-T-1216 (1216), 15 nM SB-T-121605 (121605), 5 nM SB-T-121606 (121606) or fresh medium without taxanes (CTRL) for 24 and 72 h. Expression of P-Bcl-2 (Ser70), PARP, and cleaved caspase-3 was analyzed employing western blot and relevant antibodies. Actin was used as a loading control. Data of one representative experiment out of three independent experiments shown. (B) Densitometric analysis of P-Bcl-2 (Ser70), cleaved form of PARP, and cleaved caspase-3. Data expressed as fold change of the respective control cells after 24 h and 72 h of treatment. The mean ± SD of three independent experiments shown. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: BxPC3 , Cell Lines Service GmbH Company, Eppelheim, Baden- Dr.-Eckener-Str. 8, Eppelheim, Baden-Württemberg, 69214, Germany , Product number: 305031.

Techniques: Western Blot, Expressing, Control

Journal: iScience

Article Title: Third-generation taxanes SB-T-121605 and SB-T-121606 are effective in pancreatic ductal adenocarcinoma

doi: 10.1016/j.isci.2024.109044

Figure Lengend Snippet: The analysis of cell death (A) Real-time monitoring by SYTOX Green with concentrations corresponding to IC 50 values (5–30 nM differing for each taxane-cell line combination), 100 and 300 nM PTX and SB-Ts. The data represent mean ± SD of three independent measurements. (B) The analysis of cell death by Annexin V and propidium iodide after 24, 48, and 72 h of incubation of cells with IC 50 and 100 nM concentrations of each taxane. All data presented as mean ± standard deviation. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. (C) Caspase 3 and 7 activity after 24 and 72 h of Paca-44 and BxPC-3 cells incubation with PXT and SB-Ts. Data are displayed as % of caspase activity in control cells incubated with cultivation medium only. All data presented as mean ± standard deviation. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: BxPC3 , Cell Lines Service GmbH Company, Eppelheim, Baden- Dr.-Eckener-Str. 8, Eppelheim, Baden-Württemberg, 69214, Germany , Product number: 305031.

Techniques: Incubation, Standard Deviation, Activity Assay, Control

Journal: iScience

Article Title: Third-generation taxanes SB-T-121605 and SB-T-121606 are effective in pancreatic ductal adenocarcinoma

doi: 10.1016/j.isci.2024.109044

Figure Lengend Snippet:

Article Snippet: BxPC3 , Cell Lines Service GmbH Company, Eppelheim, Baden- Dr.-Eckener-Str. 8, Eppelheim, Baden-Württemberg, 69214, Germany , Product number: 305031.

Techniques: Recombinant

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Ex vivo PD-L1/PD-1 pathway blockade reverses dysfunction of circulating CEA specific T cells in pancreatic cancer patients

doi: 10.1158/1078-0432.CCR-17-1185

Figure Lengend Snippet: (A) Pie chart illustrates the frequency of pancreatic cancer patients at different disease stages (II–IV). Frequencies of IFN-γ+ cells within the CD8+ T-cell population are shown for patients stratified according to (B) disease stage, (C) prior administration of chemotherapy, and (D) submission to surgery. Each symbol represents one individual and horizontal bars represent median. P values <0.5 were considered statistically significant and are shown in the graphs.

Article Snippet: Cell lines Six pancreatic cancer cell lines (MiaPaca-2, PK-45, Panc-1, KLM-1, Bx-Pc-3, and PK-1) were obtained from PIKEN BioResource centre (PIKEN BRC, Tsukuba, Japan).

Techniques:

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Ex vivo PD-L1/PD-1 pathway blockade reverses dysfunction of circulating CEA specific T cells in pancreatic cancer patients

doi: 10.1158/1078-0432.CCR-17-1185

Figure Lengend Snippet: (A) CEA and HLA-A2 expression of 6 pancreatic cancer cell lines. The results were analysed by FACS and presented in histogram (upper). Isotype antibodies were used to determine the background. Percentage and MFI of HLA-A2/CEA expressions by different PC cell lines was also shown (bottom). N.B. The unstained cells were gated out. (B) FACS analysis of CA11 T cells killing activity in response to recognise and kill PK-1 cell line (HLA-A2+, CEA+, labelled with high dose CFSE), and MiaPaca-2 cell line (HLA-A2−, CEA−, labelled with low dose CFSE), at different E:T ratios. (C) The percentage of relative killing of PK-1, Panc-1 and Bx-Pc-3 by CTLs from CA07 or CA11, compared to MiaPaca-2, at different E:T ratios. All the experiments were repeated twice and the mean of the results was shown in the figure.

Article Snippet: Cell lines Six pancreatic cancer cell lines (MiaPaca-2, PK-45, Panc-1, KLM-1, Bx-Pc-3, and PK-1) were obtained from PIKEN BioResource centre (PIKEN BRC, Tsukuba, Japan).

Techniques: Expressing, Activity Assay

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Ex vivo PD-L1/PD-1 pathway blockade reverses dysfunction of circulating CEA specific T cells in pancreatic cancer patients

doi: 10.1158/1078-0432.CCR-17-1185

Figure Lengend Snippet: PBMC were isolated from pancreatic cancer (PC) patients and healthy controls, and surface stained to assess (A) the expression of PD1, TIM-3 and LAG-3 molecules; as well as (B) the relative proportions of naïve/memory subsets (based on the expression of CD45RO and CD62L), within the CD8+ T-cell population. Pancreatic cancer patients were stratified according to their ability to mount CEA691-specific CD8+ T-cell responses (responders vs. non-responders). Each symbol represents one individual and horizontal bars represent mean. P values <0.5 were considered statistically significant and are shown in the graphs. (C)T cells were obtained from matching samples of peripheral blood and lymph nodes from three pancreatic cancer patients. Graphs illustrate the levels of negative regulatory molecule expression within the CD8+ population, both in terms of frequency and number of molecules per cell, expressed as mean fluorescence intensity (MFI). Each symbol represents one individual and horizontal bars represent mean. P values <0.5 were considered statistically significant and are shown in the graph.

Article Snippet: Cell lines Six pancreatic cancer cell lines (MiaPaca-2, PK-45, Panc-1, KLM-1, Bx-Pc-3, and PK-1) were obtained from PIKEN BioResource centre (PIKEN BRC, Tsukuba, Japan).

Techniques: Isolation, Staining, Expressing, Fluorescence

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Ex vivo PD-L1/PD-1 pathway blockade reverses dysfunction of circulating CEA specific T cells in pancreatic cancer patients

doi: 10.1158/1078-0432.CCR-17-1185

Figure Lengend Snippet: (A) The percentage of CEA tetramer binding CD8+ T cells after seven days of culture with CEA691 in the presence or absence of anti-PD-L1 and/or anti-TIM-3 blocking antibodies. (B) CEA691 CD8+ T cells from 11 pancreatic cancer (PC) patients were expanded for seven days with or without anti-PD-L1 and/or anti-TIM-3 blocking antibodies, and intracellular stained to assess the levels of IFN-γ production. The graph shows frequencies of IFN-γ+ cells within the CD8+ T-cell population. Each symbol represents one individual and horizontal bars represent mean. P values <0.5 were considered statistically significant and are shown in the graph. PBMCs and MNCs isolated from lymph nodes of (CA13) were also stimulated for seven days by pCEA691 with or without anti-PD-L1 and/or anti-TIM-3 blocking antibodies, and assessed for levels of IFN-γ production. A representative example of IFN-γ production is shown in the dot-plots (C). (D) The graph shows the frequency of IFN-γ+ cells within the CD8+ T-cell population. Experiments are duplicated and symbols indicate mean and SD.

Article Snippet: Cell lines Six pancreatic cancer cell lines (MiaPaca-2, PK-45, Panc-1, KLM-1, Bx-Pc-3, and PK-1) were obtained from PIKEN BioResource centre (PIKEN BRC, Tsukuba, Japan).

Techniques: Binding Assay, Blocking Assay, Staining, Isolation

Journal: International Journal of Bioprinting

Article Title: Bioprinting of hydrogel beads to engineer pancreatic tumor-stroma microtissues for drug screening

doi: 10.18063/ijb.676

Figure Lengend Snippet: Schematic presentation of this study. Three-dimensional pancreatic ductal adenocarcinoma (PDAC) models were fabricated by bioprinting of pancreatic cancer BxPC-3 cells and normal human dermal fibroblast cells laden-GelMA hydrogel beads using the dot extrusion printing technology. Drug treatment on the uniform-sized PDAC models was conducted after 1-week culture.

Article Snippet: Human pancreatic cancer cell line (BxPC-3) and normal human dermal fibroblast cells (NHDFs) in this work were kindly provided by Hangzhou Regenovo Biotechnology Co., Ltd. For cells cultivation, BxPC-3 and NHDFs were cultured in RPMI-1640 and Dulbecco’s modified Eagle’s medium (DMEM; Gibco, USA), respectively.

Techniques:

Journal: International Journal of Bioprinting

Article Title: Bioprinting of hydrogel beads to engineer pancreatic tumor-stroma microtissues for drug screening

doi: 10.18063/ijb.676

Figure Lengend Snippet: Cell viability analysis of the engineered three-dimensional pancreatic ductal adenocarcinoma (PDAC) models. (A) Representative fluorescent micrographs of three different PDAC models including mono-tumor, stroma-poor and stroma-rich microtissues at days 1, 4, and 7. Scale bar = 200 μm. (B) Cell viability. Experimental values are expressed in mean ± standard error, n = 3.

Article Snippet: Human pancreatic cancer cell line (BxPC-3) and normal human dermal fibroblast cells (NHDFs) in this work were kindly provided by Hangzhou Regenovo Biotechnology Co., Ltd. For cells cultivation, BxPC-3 and NHDFs were cultured in RPMI-1640 and Dulbecco’s modified Eagle’s medium (DMEM; Gibco, USA), respectively.

Techniques:

Journal: International Journal of Bioprinting

Article Title: Bioprinting of hydrogel beads to engineer pancreatic tumor-stroma microtissues for drug screening

doi: 10.18063/ijb.676

Figure Lengend Snippet: Morphology and organization of cells embedded in GelMA hydrogel beads of different pancreatic ductal adenocarcinoma (PDAC) models during 1-week culture. (A) F-actin staining of cell morphology and structure at day 1, 4 and 7. Green channel: F-actin. Blue channel: DAPI. Scale bar = 100 μm. (B) Assessment of the proliferation of different PDAC microtissues. (C) Assessment of the density of tumor microstructure. Experimental values are expressed in mean ± standard error, n = 3. Two-way ANOVA, * P < 0.05, *** P < 0.001, **** P < 0.0001.

Article Snippet: Human pancreatic cancer cell line (BxPC-3) and normal human dermal fibroblast cells (NHDFs) in this work were kindly provided by Hangzhou Regenovo Biotechnology Co., Ltd. For cells cultivation, BxPC-3 and NHDFs were cultured in RPMI-1640 and Dulbecco’s modified Eagle’s medium (DMEM; Gibco, USA), respectively.

Techniques: Staining

Journal: International Journal of Bioprinting

Article Title: Bioprinting of hydrogel beads to engineer pancreatic tumor-stroma microtissues for drug screening

doi: 10.18063/ijb.676

Figure Lengend Snippet: Tunable tumor-stroma microenvironment profiling. (A) Immunostaining of BxPC-3 cells and normal human dermal fibroblast cells with CK19 and α-SMA in different pancreatic ductal adenocarcinoma (PDAC) models at day 1, and 7. Green channel: α-SMA. Red channel: CK19. Blue channel: DAPI. Scale bar = 200 μm. (B) Quantitation of α-SMA expression in the co-culture models. (C) Quantitation of CK19 expression in different PDAC models. Experimental values are expressed in mean ± standard error, n = 3. Two-way ANOVA, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Human pancreatic cancer cell line (BxPC-3) and normal human dermal fibroblast cells (NHDFs) in this work were kindly provided by Hangzhou Regenovo Biotechnology Co., Ltd. For cells cultivation, BxPC-3 and NHDFs were cultured in RPMI-1640 and Dulbecco’s modified Eagle’s medium (DMEM; Gibco, USA), respectively.

Techniques: Immunostaining, Quantitation Assay, Expressing, Co-Culture Assay

Journal: International Journal of Bioprinting

Article Title: Bioprinting of hydrogel beads to engineer pancreatic tumor-stroma microtissues for drug screening

doi: 10.18063/ijb.676

Figure Lengend Snippet: Drugs screening performed in different three-dimensional pancreatic ductal adenocarcinoma (PDAC) models after 7 days of culture. (A) Representative fluorescent micrographs of PDAC models after incubation with gemcitabine ranging from 50 to 100 μM/mL for 72 h. Non-drug-treated microtissues were observed as controls. Green channel: Calcein-AM. Red channel: PI. Scale bar = 200 μm. (B) Statistical plot of cell viabilities after treating PDAC models with different concentrations of gemcitabine for 72 h. (C) Heat map representing the drug treatment results. Experimental values are expressed in mean ± standard error, n = 3. Two-way ANOVA, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Human pancreatic cancer cell line (BxPC-3) and normal human dermal fibroblast cells (NHDFs) in this work were kindly provided by Hangzhou Regenovo Biotechnology Co., Ltd. For cells cultivation, BxPC-3 and NHDFs were cultured in RPMI-1640 and Dulbecco’s modified Eagle’s medium (DMEM; Gibco, USA), respectively.

Techniques: Incubation