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: Paca-44 (carrying KRAS G12V and TP53 C176S mutations) was obtained from Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany (product. no. ACC 179) and the BxPC-3 cell line ( KRAS wild type) was obtained from CLS (Eppelheim, Germany, product no. 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: Paca-44 (carrying KRAS G12V and TP53 C176S mutations) was obtained from Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany (product. no. ACC 179) and the BxPC-3 cell line ( KRAS wild type) was obtained from CLS (Eppelheim, Germany, product no. 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: Paca-44 (carrying KRAS G12V and TP53 C176S mutations) was obtained from Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany (product. no. ACC 179) and the BxPC-3 cell line ( KRAS wild type) was obtained from CLS (Eppelheim, Germany, product no. 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: Paca-44 (carrying KRAS G12V and TP53 C176S mutations) was obtained from Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany (product. no. ACC 179) and the BxPC-3 cell line ( KRAS wild type) was obtained from CLS (Eppelheim, Germany, product no. 305031).

Techniques: Incubation, Standard Deviation, Activity Assay, Control

Journal: Theranostics

Article Title: MSC-Delivered Soluble TRAIL and Paclitaxel as Novel Combinatory Treatment for Pancreatic Adenocarcinoma.

doi: 10.7150/thno.27576

Figure Lengend Snippet: Figure 1. Generation of BXPC3 clones resistant to sTRAIL and rhTRAIL. (A) Representation of the selection strategy. Wild type BXPC3 were treated with sTRAIL- containing supernatant once a week for 10 weeks (pulsed selection), obtaining PA and PB clones, or constantly for 10 weeks (continuous selection), obtaining CA and CB clones. (B) Wild type BXPC3 and selected clones were tested for susceptibility to sTRAIL treatment by MTS assay. Each bar represents metabolic inhibition respect to untreated control. (C) rhTRAIL dose-response assay for wild type and selected BXPC3 clones in MTS. Cells were treated with a range of concentrations of rhTRAIL for 24 h. (D) Expression of TRAIL functional (DR4, DR5) and decoy (DcR1, DcR2) receptors by flow cytometry on wild type and selected BXPC3 clones. (E) qPCR analysis on PA clone for the expression of key pro-survival genes respect to BXPC3 wild type cells. Data are represented as mean±SD. All data are represented as the mean of two independent experiments performed in triplicate. WT: Wild type BXPC3; CA, CB, PA, PB: selected BXPC3 clones. * p<0.01, ** p<0.001, *** p<0.0001 with Student’s T-test. § p<0.05, ^ p<0.0001, ° p<0.0001 by ANOVA and multiple comparison test.

Article Snippet: Authentication of BXPC3 cells was performed last year by Leibniz-Institut DSMZ GmbH (Braunschweig, Germany).

Techniques: Clone Assay, Selection, MTS Assay, Inhibition, Control, Expressing, Functional Assay, Flow Cytometry, Comparison

Journal: Theranostics

Article Title: MSC-Delivered Soluble TRAIL and Paclitaxel as Novel Combinatory Treatment for Pancreatic Adenocarcinoma.

doi: 10.7150/thno.27576

Figure Lengend Snippet: Figure 2. Paclitaxel treatment sensitizes resistant BXPC3 to sTRAIL by reversing survival pathways (A) PTX dose-response assay for wild type (WT) and selected BXPC3 clones by MTS. Cells were treated with a range of concentrations of PTX for 72 h. (B) In vitro combined treatment of sTRAIL- containing supernatant with PTX. PA clone cells were treated for 24 h with AD-MSC EMPTY derived conditioned medium (EMPTY CM) or AD-MSC sTRAIL derived conditioned medium (sTRAIL CM) or the same with 5 ng/mL PTX (EMPTY CM+PTX and sTRAIL CM+PTX). Death rate was evaluated by PI and Annexin V staining in flow cytometry. (C) Pre-treatment strategy included a 24 h incubation with 5 ng/mL PTX for EMPTY CM+PTX and sTRAIL CM+PTX groups. The subsequent treatment was performed as in B for a further 24 h. (D) Expression of main pro-survival genes by qPCR assay on BXPC3 PA after 24 h treatment with 50 ng/mL PTX (BXPC3 PA-PTX) compared with untreated cells (BXPC3 PA) and BXPC3 WT as baseline. Data are represented as mean±SD. All data are represented as the mean of two independent experiments performed in triplicate. * p<0.01; ** p<0.05 with Student’s T-test.

Article Snippet: Authentication of BXPC3 cells was performed last year by Leibniz-Institut DSMZ GmbH (Braunschweig, Germany).

Techniques: Clone Assay, In Vitro, Derivative Assay, Staining, Flow Cytometry, Incubation, Expressing

Journal: Theranostics

Article Title: MSC-Delivered Soluble TRAIL and Paclitaxel as Novel Combinatory Treatment for Pancreatic Adenocarcinoma.

doi: 10.7150/thno.27576

Figure Lengend Snippet: Figure 3. Inhibition of NFkB confirms its main role in sTRAIL resistance of selected BXPC3. (A) DHMEQ dose-response assay for wild type (WT) and selected BXPC3 clones by MTS. Cells were treated with a range of concentrations of DHMEQ for 48 h. (B) In vitro combined treatment of sTRAIL-containing supernatant with DHMEQ. PA clone cells were treated for 24 h with AD-MSC EMPTY derived conditioned medium (EMPTY CM) or AD-MSC sTRAIL derived conditioned medium (sTRAIL CM) or the same with 20 µg/mL DHMEQ (EMPTY CM+DHMEQ and sTRAIL CM+DHMEQ). Death rate was evaluated by PI and Annexin V staining in flow cytometry. All data are represented as the mean of two independent experiments performed in triplicate. * p<0.05; ** p<0.01; *** p<0.001; **** p<0.0001 with Student’s T-test

Article Snippet: Authentication of BXPC3 cells was performed last year by Leibniz-Institut DSMZ GmbH (Braunschweig, Germany).

Techniques: Inhibition, Clone Assay, In Vitro, Derivative Assay, Staining, Flow Cytometry

Journal: Journal of colloid and interface science

Article Title: Engineering magnetic nanoparticles and their integration with microfluidics for cell isolation

doi: 10.1016/j.jcis.2019.12.092

Figure Lengend Snippet: Uptake of iron oxide nanoparticles in BxPC3 and CCRF-CEM cells for cells with no particles (in red), cells incubated with nontargeted particles (in blue), and cells incubated with targeted particles (in green). All experiments were repeated 3 times.

Article Snippet: In contrast, the purity of captured cells was close to 100% for the BxPC3 cells mixed with CEM cells in PBS, while it was about 34% for the BxPC3 cells in whole blood.

Techniques: Incubation

Journal: Journal of colloid and interface science

Article Title: Engineering magnetic nanoparticles and their integration with microfluidics for cell isolation

doi: 10.1016/j.jcis.2019.12.092

Figure Lengend Snippet: Interaction of targeted (top panel) and non-targeted nanoparticles (bottom panel) with BxPC3 cells.

Article Snippet: In contrast, the purity of captured cells was close to 100% for the BxPC3 cells mixed with CEM cells in PBS, while it was about 34% for the BxPC3 cells in whole blood.

Techniques:

Journal: Journal of colloid and interface science

Article Title: Engineering magnetic nanoparticles and their integration with microfluidics for cell isolation

doi: 10.1016/j.jcis.2019.12.092

Figure Lengend Snippet: BxPC3 cells with targeted nanoparticles indicated by white arrows.

Article Snippet: In contrast, the purity of captured cells was close to 100% for the BxPC3 cells mixed with CEM cells in PBS, while it was about 34% for the BxPC3 cells in whole blood.

Techniques:

Journal: Journal of colloid and interface science

Article Title: Engineering magnetic nanoparticles and their integration with microfluidics for cell isolation

doi: 10.1016/j.jcis.2019.12.092

Figure Lengend Snippet: Capture efficiencies of BxPC3 (Red) and CCRF-CEM (blue) cells with (solid bar) and without (dashed bar) application of the Halbach array. All experiments were conducted in 3 different microfluidic devices.

Article Snippet: In contrast, the purity of captured cells was close to 100% for the BxPC3 cells mixed with CEM cells in PBS, while it was about 34% for the BxPC3 cells in whole blood.

Techniques:

Journal: Journal of colloid and interface science

Article Title: Engineering magnetic nanoparticles and their integration with microfluidics for cell isolation

doi: 10.1016/j.jcis.2019.12.092

Figure Lengend Snippet: a. Capture efficiency of BxPC3 when spiked in blood mixture in red and the purity of capture in grey; b. Representative image of the BxPC3 cells (membrane stained with cell mask deep red, nuclei stained with Hoescht 333422) captured against white blood cells (Nuclei stained with Hoescht 333422) in whole blood. All experiments were conducted in 3 different microfluidic devices.

Article Snippet: In contrast, the purity of captured cells was close to 100% for the BxPC3 cells mixed with CEM cells in PBS, while it was about 34% for the BxPC3 cells in whole blood.

Techniques: Membrane, Staining