breast cancer cell lines mda mb 231  (ATCC)

Journal: Bioactive Materials

Article Title: Ribbon-shaped microgels as bioinks for 3D bioprinting of anisotropic tissue structures

doi: 10.1016/j.bioactmat.2025.12.040

Figure Lengend Snippet: μRB bioinks support multicellular patterning to model breast cancer-bone invasion at tissue interface. (A) Schematic of experimental design: bioprinting of MSCs and osteogenic differentiation to derive bone grid, followed by injection of breast cancer bioink, and monitoring invasion over time using confocal microscopy. (B) Confocal images of MSCs (red) after printing (Scale bar = 1 mm). (C) Confocal images of scaffold sections containing CellTracker-labeled MSCs (red) after 28 days of osteogenic differentiation and GFP + MDA-MB-231 cells extruded into the open pores of the grids (green) (Scale bar = 200 μm). (D) Confocal images of patterned MSC-derived bone (red) with MDA-MB-231 and MCF-7 breast cancer cells (green) after 14 days of co-culture (Scale bar = 1 mm). (E) Quantification of breast cancer cell invasion: percentage that remain in open pores vs. invading into the MSC-bone compartment (n = 5 per group). Values are reported as mean ± S.D. and p-values were determined by two-way analysis of variance (ANOVA) with Tukey's multiple comparisons test; ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.005, ∗∗∗∗p ≤ 0.001.

Article Snippet: Breast cancer cell lines MDA-MB-231 (ATCC) and MCF-7 (ATCC) were lentivirus transduced to express GFP and cultured in DMEM media (4.5 g L −1 glucose) supplemented with 10 % (v/v) fetal bovine serum and 1 % (v/v) penicillin-streptomycin.

Techniques: Injection, Confocal Microscopy, Labeling, Derivative Assay, Co-Culture Assay

Journal: International Journal of Molecular Medicine

Article Title: Ebastine targets HER2/HER3 signaling and cancer stem cell traits to overcome trastuzumab resistance in HER2-positive breast cancer

doi: 10.3892/ijmm.2026.5751

Figure Lengend Snippet: EBA impairs cancer stem cell-like properties. (A) BT474 and SKBR3 cells were treated with EBA for 48 h, and ALDH1 activity was assessed by flow cytometry using the Aldefluor assay. DEAB was used to define the baseline of Aldefluor-positive fluorescence. (B) BT474 cells (5x10 4 cells/ml) were plated in ultra-low attachment dishes and cultured in the presence or absence of EBA for 5 days. The number and volume of mammospheres were measured by microscopy. (C) Overall survival of patients with breast cancer stratified by the co-expression of ALDH1A1 and CD44. (D) Spearman correlation analysis of ALDH1A1 and CD44 mRNA levels in patients with HER2-positive breast cancer from The Cancer Genome Atlas cohort (n=76). Kaplan-Meier survival analyses of patients with HER2-overexpressing breast cancer stratified by (E) ALDH1A1 and (F) CD44 expression. Patients were divided into high- and low-expression groups based on the median gene expression. Statistical significance was determined using the log-rank test. (G) JIMT-1 cells were treated with EBA (3 μ M) for 48 h and the CD44 high /CD24 low cell populations were identified by flow cytometry. (H) JIMT-1 cells (1.5x10 4 cells/ml) were cultured under serum-free suspension conditions in the presence of EBA (3 μ M) for 8 days. Mammosphere number and volumes were quantified. ** P<0.01 and **** P<0.0001 vs. vehicle-treated control (0 μ M EBA). EBA, ebastine; ALDH, aldehyde dehydrogenase; DEAB, diethylaminobenzaldehyde; CTL, control; ISO, isotype.

Article Snippet: The human breast cancer cell lines SKBR3, BT474, MDA-MB-453 (American Type Culture Collection) and JIMT-1 (Leibnitz Institute DSMZ-German Collection of Microorganisms and Cell Cultures GmbH) were cultured in DMEM, MEM or RPMI-1640 (all Sigma-Aldrich; Merck KGaA) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) and 100 U/ml penicillin-streptomycin at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Activity Assay, Flow Cytometry, Fluorescence, Cell Culture, Microscopy, Expressing, Gene Expression, Suspension, Control

Journal: bioRxiv

Article Title: Single-cell profiling of synchronous multi-organ metastasis reveals a systemic CD74 + lipid-associated macrophage niche driving polymetastatic breast cancer

doi: 10.64898/2026.01.31.701004

Figure Lengend Snippet: a. qPCR analysis of Mif expression in VO-PyMT cells transduced with control or two independent Mif-targeting short hairpin RNAs (shRNA). Bar heights show means and error bars depict standard deviation of technical triplicates. Data was reproduced in 3 independent experiments. b. Immunoblot of Mif protein in shControl- or shMif-transduced VO-PyMT cells. B-Actin is shown as loading control. Data is representative of two independent experiments. c. MIF protein concentration in cell culture supernatants of shControl and shMif-transduced VO-PyMT cells as determined by enzyme linked immunosorbent assay (ELISA). Bar heights depict means and error bars show standard deviation of two technical replicates. d. Relative Mif expression levels in Py8119 cells harboring an shControl or shMif hairpin, as determined by qPCR. Error bars show standard deviations. Data is representative of 3 separate experiments. e. Western blot analysis of MIF protein in shControl or shMif Py8119 cells. B-Actin is shown as loading control. Data is representative of two independent experiments. f. Number of spheres per well quantified in in vitro spheroid cultures of VO-PyMT or Py8119 cells upon Mif knockdown. Sphere count was performed 7 days post seeding. Ns, not significant. Two-tailed Student’s t tests were used to estimate significance. g. Representative images of spheroids quantified in (f). Scale bars: 1 mm.

Article Snippet: Py8119 mouse breast cancer cells were purchased from ATCC (cat. no. CRL-3278), and cultured in F-12K medium supplemented with 2 mM L-Glutamine, 1,500 mg/L sodium bicarbonate, 10% vol/vol FBS, and P/S.

Techniques: Expressing, Transduction, Control, shRNA, Standard Deviation, Western Blot, Protein Concentration, Cell Culture, Enzyme-linked Immunosorbent Assay, In Vitro, Knockdown, Two Tailed Test

Journal: bioRxiv

Article Title: Single-cell profiling of synchronous multi-organ metastasis reveals a systemic CD74 + lipid-associated macrophage niche driving polymetastatic breast cancer

doi: 10.64898/2026.01.31.701004

Figure Lengend Snippet: a. Quantification of metastatic burden using ex vivo BLI of FVB mice injected VO-PyMT cells transduced with control vectors (shControl) and Mif-knockdown vectors (shMif 1; shMif 2). Data was acquired at day 12 post i.c. injections. shControl, n = 11; shMif (1), n = 11; shMif (2), n = 5. Data represents results from 2 independent experiments. In each experiment, raw photon flux values (p/s) from each organ were normalized to the average photon flux (p/s) values of the shControl group. Boxes depict 25th and 75th percentiles. Median is shown as horizontal lines. Whiskers depict data range. All data points are shown as dots. P values were determined using two-tailed Mann-Whitney tests by comparing shControl ( n = 11) versus the two shMif groups combined ( n = 16). Images show representative ex vivo BLI in brain, lung, liver or lower limb bones from each experimental group. b. Quantification of multi-organ metastatic burden ex vivo using BLI in albino C57/BL6 mice injected intracardially with Py8119 breast cancer cells transduced with control (shControl) and Mif-knockdown vector (shMif 2). Representative images are shown for each organ analyzed. Ex vivo metastatic burden was quantified 10 days after cancer cell injection. shControl, n = 14; shMif (2), n = 14. Data is representative of 3 independent experiments. Data in each organ was normalized to the average photon flux (p/s) of the shControl group in each experiment. Boxes boundaries define the interquartile ranges. Horizontal lines depict median values in each group. The whiskers show data range. The dots depict data points. P values were calculated by one-tailed Mann-Whitney t tests. c. Schematic showing pre-clinical model of MIF-CD74 targeting using 4-IPP, a small molecule inhibitor of MIF binding capacity. Multi-metastatic VO-PyMT cells were injected intracardially at day 0. Seeding and micrometastatic growth was allowed for 3 days, followed by i.p. administration of either a vehicle solution or 4-IPP. At day 12 post i.c. injection of VO-PyMT cells, metastatic colonization was analyzed ex vivo in brain, lung, liver and lower limb bones. d. Box plots showing quantification of metastatic burden of experiment as determined by ex vivo BLI in brain, lung, liver and bones of FVB mice injected intracardially with VO-PyMT cells and treated with the MIF inhibitor 4-IPP as described in (c). Vehicle group, n = 14; 4-IPP group, n = 13. For bone metastatic burden, right and left lower limb bones were analyzed separately. Data represents results from 2 independent experiments. In each experiment, photon flux (p/s) in each organ was normalized to the average photon flux (p/s) in the Vehicle control group. Boxes boundaries indicate 25 th and 75 th percentiles. Median is indicated by a horizontal line in boxes. Whiskers show data range. Data points are indicated as dots. P values were calculated using one-tailed Mann-Whitney t tests.

Article Snippet: Py8119 mouse breast cancer cells were purchased from ATCC (cat. no. CRL-3278), and cultured in F-12K medium supplemented with 2 mM L-Glutamine, 1,500 mg/L sodium bicarbonate, 10% vol/vol FBS, and P/S.

Techniques: Ex Vivo, Injection, Transduction, Control, Knockdown, Two Tailed Test, MANN-WHITNEY, Plasmid Preparation, One-tailed Test, Binding Assay