Journal: STAR Protocols

Article Title: Protocol for enhancing CRISPR-Cas9 genome editing using histone deacetylase inhibition and engineered virus-like particle delivery

doi: 10.1016/j.xpro.2026.104493

Figure Lengend Snippet: FAME-CRISPR improves CRISPR/Cas9 gene editing efficiency through HDACi mediated chromatin relaxation and precise eVLP delivery (A) shows a brief schematic of the major steps required to improve editing efficacy using FAME-CRISPR workflow. (B), (C), (D), and (E) show representative results obtained from MDA-MB-231 one of the utilized cell lines from the original FAME-CRISPR study ([60nM panobinostat] and [1.5 μM belinostat]) (n = 3 biological replicates for all conditions, and error bars reflect standard deviation). Results and raw data for MDA-MB-231 and other tested cell lines are available in the original study and data availability statement section of STAR Protocols.

Article Snippet: MDA-MB-231 , ATCC , #HTB-26.

Techniques: CRISPR, Standard Deviation

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: Ptch1 is expressed in breast cancer cell lines. A. PTCH1 mRNA expression (log 2 transformed) in various luminal and HER2 breast cancer cell lines (dark green) and in TNBC cell lines (other colors). TNBC cell lines are depicted according to the “Lehmann TNBC subtype” nomenclature : basal-like 1 (yellow), basal-like 2 (pale green), immunomodulatory (brown), luminal androgen receptor (dark pink), mesenchymal (pale pink) and mesenchymal stem-like (pink). B. PTCH1 protein expression in three TNBC cell lines. Western-blot was performed on 50 µg extracts from TNBC cell lines (MDA-MB-231, HCC-38 and MDA-MB-468) with antibodies directed against PTCH1. PTCH1 and GAPDH signals were quantified using ImageJ software. Data presented are the mean ± SEM of at least 3 independent experiments. Significance is calculated using Oneway ANOVA Turkey’s multiple comparisons test and attained at P < 0.05 (*).

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Expressing, Transformation Assay, Western Blot, Software

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: PTCH1 drug efflux inhibitor PAH increases the sensitivity of TNBC cells to chemotherapy . Cell viability was measured after 24 h or 48 h treatment with increasing concentration of docetaxel or doxorubicin respectively on MDA-MB-231, MDA-MB-468 and HCC-38 cell lines in the absence or the presence of 15µM PAH. IC 50 values (corresponding to the concentration of chemotherapy inducing 50% of cell death) were calculated. Data reported are the mean ± SEM of at least 3 independent experiments. Significance is attained at P < 0.05 (*).

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Concentration Assay

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: Other multidrug transporters are expressed in TNBC cell lines. A. P-gp and ABCG2 protein expression in three TNBC cell lines. Western-blot was performed on 50 µg extracts from each TNBC cell line (MDA-MB-231, HCC-38 and MDA-MB-468) with antibodies directed against P-gp or ABCG2 and GAPDH. B. P-gp, ABCG2 and GAPDH signals were quantified using ImageJ software. Data presented are the mean ± SEM of at least 3 independent experiments. Significance is calculated using Oneway ANOVA Turkey’s multiple comparisons test and attained at P < 0.05 (*). ** P < 0.01; *** P < 0.001; ns P > 0.05.

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Expressing, Western Blot, Software

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: PAH increases docetaxel efficacy against TNBC spheroids. MDA-MB-231 cells were plated in ultra-low attachment surface 24 well plates in complete medium and treated with increasing concentrations of docetaxel in the presence of DMSO (control, 0 PAH), PAH 10 µM or 30 µM. After 2 weeks pictures were taken using Cytation 5 cell imaging system from Biotek. The surface of spheroids was calculated and reported after normalization on the condition without docetaxel for each concentration of PAH.

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Control, Imaging, Concentration Assay

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: PAH addition to chemotherapy inhibits migration of TNBC cells. A. 100000 cells were seeded on membrane from Transwell plates. Cells were treated 24 h with doxorubicin ± 15µM PAH or 48 h with docetaxel ± 15µM PAH. After fixation and staining with crystal violet, wells were observed on a microscope with X5 objective. B. Migration was measured using a wound-healing assay. A wound was performed on MDA-MB-231 confluent cells seeded in 24 well plates. The medium was replaced by fresh medium containing 5 µM docetaxel in the absence of PAH, or the presence of 5 µM PAH. Two pictures were taken at two different points of each well immediately after wound, and 3 days after wound with 5X objective. The width of the wound was measured using ImageJ software and reported as final wound width/initial wound width in percentage. Data presented are the mean ± SEM of 3 independent experiments. Significance is attained at P < 0.05.

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Migration, Membrane, Staining, Microscopy, Wound Healing Assay, Software

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: PTCH1 contributes to the efflux of doxorubicin from TNBC cells. A. PTCH1 protein expression (left panel) and intracellular doxorubicin (right panel) were analyzed 16 h after transfection of MDA-MB-231 cells with PTCH1-siRNA or negative-control-siRNA. PTCH1 and GAPDH western blot signals were quantified using ImageJ software (left panel). For doxorubicin accumulation measurements (right panel), MDA-MB-231 cells were grown on slides and transfected with PTCH1-siRNA or negative-control-siRNA. After incubation with 10 µM doxorubicin, 3 coverslips were fixed for doxorubicin loading control; the other coverslips were incubated with efflux buffer and fixed. B. Docetaxel inhibits the accumulation of doxorubicin in MDA-MB-231 cells. Cells on coverslip were incubated with 10 µM doxorubicin or 10 µM doxorubicin and 50 µM docetaxel. Doxorubicin fluorescence images were acquired by epifluorescence microscopy using a 40X objective, and doxorubicin fluorescence was quantified using ImageJ software for about 100 cells per condition per experiment. Histograms are the mean ± SEM of 3 independent experiments. Significance is attained at P < 0.05.

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Expressing, Transfection, Negative Control, Western Blot, Software, Incubation, Control, Fluorescence, Epifluorescence Microscopy

Journal: Materials Today Bio

Article Title: Mannosylated graphene oxide nanotherapeutics co-delivering docetaxel and a STING agonist reprogram myeloid cells and potentiate antitumor immunity

doi: 10.1016/j.mtbio.2026.103086

Figure Lengend Snippet: Macrophage-targeted GO-EDM-DTX-Vad induces M2-to-M1 repolarization and limits TNBC cell proliferation and migration. (A) After incubation of M2-polarized macrophages with the nanocomposites for 24 h, CD206 and CD86 were quantified by flow cytometry. (B) After co-treatment of M0 RAW264.7 cells with IL-4/IL-13 and the nanocomposites for 48 h, CD206 and CD86 were quantified by flow cytometry. (C) RT-qPCR of CD86, iNOS, CD206 and IL-10 expression. (D) RAW264.7 immunoblots of STING axis including TBK1, phospho-TBK1, IRF3 and phospho-IRF3, with GAPDH as a reference. (E) CLSM images of RAW264.7 uptake of RhoB–GO-EDM-DTX-Vad (nuclei, blue; nanocomposites, red; scale bar, 100 μm). (F) ELISA of TGF-β, IL-10 and TNF-α in culture supernatants. (G) Schematic of the in vitro workflow for generating macrophage-conditioned media and assessing CM-driven tumor cell functions. (H, I) Colony formation (H) and EdU flow-cytometry analyses (I) of 4T1 and MDA-MB-231 cells after treatment with the indicated macrophage-CM. Groups: G1, PBS; G2, GO-EDM; G3, GO-EDM-DTX; G4, GO-EDM-Vad; G5, GO-EDM-DTX-Vad; G6, GO-EDM-DTX-Vad + NIR. Data are mean ± SD (n = 3). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001; ns, not significant.

Article Snippet: 4T1 and MDA-MB-231 cells were treated with the indicated nanocomposites for 24 h. The apoptosis was quantified by Annexin V-FITC/PI staining (Elabscience, Wuhan, China) followed by flow cytometry, and cell viability was visualized by Calcein-AM/PI live/dead staining (Servicebio, Wuhan, China) using CLSM.

Techniques: Migration, Incubation, Flow Cytometry, Quantitative RT-PCR, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, In Vitro

Journal: Materials Today Bio

Article Title: Mannosylated graphene oxide nanotherapeutics co-delivering docetaxel and a STING agonist reprogram myeloid cells and potentiate antitumor immunity

doi: 10.1016/j.mtbio.2026.103086

Figure Lengend Snippet: GO-EDM-DTX-Vad combined with NIR induces TNBC cell death, apoptosis and immunogenic cell death. (A, B) Calcein-AM/PI live/dead staining of 4T1 (A) and MDA-MB-231 (B) cells (live, green; dead, red; scale bar, 200 μm). (C, D) Annexin V/PI apoptosis plots for 4T1 (C) and MDA-MB-231 (D) cells. (E, F) Quantification of viable and early/late apoptotic 4T1 cells. (H, I) Quantification of viable and early/late apoptotic MDA-MB-231 cells. (G, J) Dose–response viability curves after 48 h treatment with free DTX or GO-EDM-DTX-Vad in 4T1 (G) and MDA-MB-231 (J). (K, L) Immunofluorescence of CRT exposure and HMGB1 release after 24 h treatment in 4T1 (K) and MDA-MB-231 (L) cells (nuclei, blue; CRT/HMGB1, green; scale bar, 100 μm). (M) Schematic of the ICD-to-DC maturation assay. (N–P) Frequencies of CD40 + , CD80 + and CD86 + DCs after co-culture. Data are mean ± SD (n = 3). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001; ns, not significant.

Article Snippet: 4T1 and MDA-MB-231 cells were treated with the indicated nanocomposites for 24 h. The apoptosis was quantified by Annexin V-FITC/PI staining (Elabscience, Wuhan, China) followed by flow cytometry, and cell viability was visualized by Calcein-AM/PI live/dead staining (Servicebio, Wuhan, China) using CLSM.

Techniques: Staining, Immunofluorescence, Co-Culture Assay

Journal: Journal of Cell Communication and Signaling

Article Title: Human brain pericytes protect the blood–brain barrier from triple‐negative breast cancer cells while promoting tumor aggressiveness

doi: 10.1002/ccs3.70070

Figure Lengend Snippet: Brain pericyte secretions mediate the inhibition of TNBC cell adhesion to the BBB. (A) Experimental workflow for adhesion assay. All BBB experiments were performed using ECGMV2 medium with supplements for all conditions from day −6 to the end of the experiment. The astrocyte medium was renewed 24 h before the adhesion experiment with the same ECGMV2 medium. Endothelial cells (ECs) and brain pericytes (hBPs) were co‐cultured for 5 days, and inserts containing brain‐like endothelial cells (hBLECs) were then either maintained with hBPs or transferred to wells without hBPs. After 24 hours, and prior to MDA‐MB‐231 cells seeding, inserts were subjected to the following conditions: maintained without hBPs (hBLEC 24h); transferred to hBPs previously cultured alone for 24 hours (hBLEC 24h then hBP); transferred to wells without hBPs but containing either culture medium (hBLEC) or CM from co‐culture (hBLEC + CM coc ); transferred to wells with astrocytes (hBLEC + A) or maintained in co‐culture with hBPs (hBLEC + hBP). (B–G), Quantification (B, D, F) of MDA‐MB‐231 cell adhesion to hBLECs after 3 h of incubation and representative images (C, E, G) of adhered MDA‐MB‐231 cells (green) under the different conditions. Data are obtained from three independent experiments, with three technical replicates per condition. Statistical analyses were performed using one‐way ANOVA followed by Tukey's test (B), Welch's ANOVA followed by Dunnett's T3 test (D) and Kruskal–Wallis test followed by Dunn's test (F). CM coc = conditioned medium from 24 h hBLECs and hBPs coculture, A = astrocytes, hBP mono = 24 h hBPs monoculture. Scale bar = 750 µm. BBB, blood–brain barrier; ECGMV2, EC Growth Medium MV 2; ECs, endothelial cells; hBLECs, human brain‐like endothelial cells; ns, non‐significant; hBPs, human brain pericytes; TNBC, triple‐negative breast cancer. **** p ≤ 0.0001; * p ≤ 0.05.

Article Snippet: The human TNBC cell line MDA‐MB‐231 (female) was obtained from the American type culture collection (ATCC, HTB‐26, RRID:CVCL_0062, obtained in 2023).

Techniques: Inhibition, Cell Adhesion Assay, Cell Culture, Co-Culture Assay, Incubation

Journal: Journal of Cell Communication and Signaling

Article Title: Human brain pericytes protect the blood–brain barrier from triple‐negative breast cancer cells while promoting tumor aggressiveness

doi: 10.1002/ccs3.70070

Figure Lengend Snippet: Brain pericytes maintain BBB integrity in the presence of TNBC cells. (A–E) Endothelial permeability (Pe) to Lucifer Yellow assessed after 3 h (A) or 16 h (C) of incubation with MDA‐MB‐231 cells, in the presence or absence of brain pericytes (hBPs). Permeability was also measured in the absence of cancer cells as a control. Permeability values (expressed in × 10 −3 cm/min ± SD) are summarized in a table (E). Representative images of endothelial Claudin‐5 (magenta) immunostaining in the absence or presence of hBPs after 3 h (B) or 16 h (D) of incubation with MDA‐MB‐231 cells (green). Data are obtained from three independent experiments, with three technical replicates per condition. Statistical analyses were performed using one‐way ANOVA followed by Tukey's test (A) and Welch's ANOVA followed by Dunnett's T3 test (B). MDA = MDA‐MB‐231 cells. Scale bars = 300 μm (10X) and 100 μm (40X). BBB, blood–brain barrier; hBPs, human brain pericytes; ns, non‐significant; TNBC, triple‐negative breast cancer. *** p ≤ 0.001; ** p ≤ 0.01; * p ≤ 0.05.

Article Snippet: The human TNBC cell line MDA‐MB‐231 (female) was obtained from the American type culture collection (ATCC, HTB‐26, RRID:CVCL_0062, obtained in 2023).

Techniques: Permeability, Incubation, Control, Immunostaining

Journal: Journal of Cell Communication and Signaling

Article Title: Human brain pericytes protect the blood–brain barrier from triple‐negative breast cancer cells while promoting tumor aggressiveness

doi: 10.1002/ccs3.70070

Figure Lengend Snippet: Brain pericytes limit TNBC cell adhesion to the BBB under hypoxic conditions. (A) Experimental workflow for adhesion assay in hypoxic conditions. All BBB experiments were performed using ECGMV2 medium with supplements for all conditions from day −6 to the end of the experiment. After 5 days of co‐culture with brain pericytes (hBPs), inserts containing brain‐like endothelial cells (hBLECs) were exposed to hypoxia for 24 h in the presence or absence of hBPs. Parallel conditions maintained in normoxia served as controls. (B–E) Quantification (B, D) of MDA‐MB‐231 cell adhesion to hBLECs after 3 h of incubation and representative images (C, E) of adhered MDA‐MB‐231 cells (green) under the different conditions. Data are obtained from three independent experiments, with three technical replicates per condition. Statistical analyses were performed using Welch's ANOVA followed by Dunnett's T3 test (B) and Kruskal‐Wallis test followed by Dunn's test (D). hBP‐CM mono = conditioned medium from 24 h hBPs monoculture. Scale bar = 750 μm. BBB, blood–brain barrier; ECGMV2, EC Growth Medium MV 2; hBLECs, human brain‐like endothelial cells; hBPs, human brain pericytes; ns, non‐significant; TNBC, triple‐negative breast cancer. *** p ≤ 0.001; ** p ≤ 0.01; * p ≤ 0.05.

Article Snippet: The human TNBC cell line MDA‐MB‐231 (female) was obtained from the American type culture collection (ATCC, HTB‐26, RRID:CVCL_0062, obtained in 2023).

Techniques: Cell Adhesion Assay, Co-Culture Assay, Incubation

Journal: Journal of Cell Communication and Signaling

Article Title: Human brain pericytes protect the blood–brain barrier from triple‐negative breast cancer cells while promoting tumor aggressiveness

doi: 10.1002/ccs3.70070

Figure Lengend Snippet: Brain pericytes maintain BBB integrity in the presence of TNBC cells under hypoxic conditions and supplement deprivation. (A) After 5 days of co‐culture with brain pericytes (hBPs) in ECGMV2 medium with supplements, inserts containing brain‐like endothelial cells (hBLECs) co‐cultured with hBPs were exposed to hypoxia for 16 h following medium replacement with ECGMV2 basal medium (without supplements). Then, endothelial permeability (Pe) to Lucifer Yellow was assessed after 3 h of incubation with MDA‐MB‐231 cells, in the presence or absence of hBPs, under normoxic and hypoxic conditions. (B) Permeability values (expressed in x10 −3 cm/min ± SD) are summarized in a table. (C) Representative images of endothelial Claudin‐5 (magenta) immunostaining in the absence or presence of hBPs after 3 h of incubation with MDA‐MB‐231 cells (green) under hypoxic conditions in basal medium. Data are obtained from three independent experiments, with three technical replicates per condition. Statistical analyses were performed using Welch's ANOVA followed by Dunnett's T3 test. MDA = MDA‐MB‐231 cells. Scale bars = 300 μm (10X) and 100 μm (40X). BBB, blood–brain barrier; ECGMV2, EC Growth Medium MV 2; hBLECs, human brain‐like endothelial cells; hBPs, human brain pericytes; ns, non‐significant; TNBC, triple‐negative breast cancer. *** p ≤ 0.001; ** p ≤ 0.01; * p ≤ 0.05.

Article Snippet: The human TNBC cell line MDA‐MB‐231 (female) was obtained from the American type culture collection (ATCC, HTB‐26, RRID:CVCL_0062, obtained in 2023).

Techniques: Co-Culture Assay, Cell Culture, Permeability, Incubation, Immunostaining

Journal: Journal of Cell Communication and Signaling

Article Title: Human brain pericytes protect the blood–brain barrier from triple‐negative breast cancer cells while promoting tumor aggressiveness

doi: 10.1002/ccs3.70070

Figure Lengend Snippet: Brain pericytes enhance migratory and invasion properties of TNBC cells. (A) Quantification of MDA‐MB‐231 cells that migrated to the lower side of insert filters in the absence (∅) or presence of brain pericytes (+hBPs), and representative images of migrated cells (nuclei stained in blue) associated. (B) Quantification of MDA‐MB‐231 cells that invaded the lower side of insert filters pre‐coated with Matrigel ® in the presence (+hBP) or absence (∅) of hBPs. (C) Representative images of MDA‐MB‐231 cells (green) after 48 h of monoculture or co‐culture with hBPs; cell morphology is visualized by F‐actin staining (phalloidin, orange). (D) Heatmap visualization of F‐actin intensity. (E) Quantification of cortical F‐actin fluorescence in MDA‐MB‐231 cells. (F) MDA‐MB‐231 cell elongation factor, defined as the ratio of cell length to width, following 48 h of monoculture (MDA) and co‐culture with hBPs (MDA + hBP), or following 1‐hour treatment with DMEM 0.1% (CTL medium) and hBP‐CM. Data are obtained from three independent experiments, with three technical replicates (A, B) or at least 30 cells analyzed (E, F) per condition. Statistical analyses were performed using a Mann–Whitney test (A, E, F) and an unpaired t ‐test with Welch's correction (B). MDA = MDA‐MB‐231 cells. Scale bars = 750 μm (A) and 100 μm (C, D). DMEM, Dulbecco'’s Modified Eagle Medium; hBP‐CM, hBP‐conditioned medium; hBPs, human brain pericytes; TNBC, triple‐negative breast cancer. **** p ≤ 0.0001; * p ≤ 0.05.

Article Snippet: The human TNBC cell line MDA‐MB‐231 (female) was obtained from the American type culture collection (ATCC, HTB‐26, RRID:CVCL_0062, obtained in 2023).

Techniques: Staining, Co-Culture Assay, Fluorescence, MANN-WHITNEY, Modification

Journal: Journal of Cell Communication and Signaling

Article Title: Human brain pericytes protect the blood–brain barrier from triple‐negative breast cancer cells while promoting tumor aggressiveness

doi: 10.1002/ccs3.70070

Figure Lengend Snippet: Brain pericyte secretions enhance TNBC clonogenic capacity. (A) Representative images of MDA‐MB‐231 colony formation following exposure to conditioned medium from brain pericytes (hBP‐CM), control medium (DMEM 0.1%, medium used for CM generation), or DMEM 10%. (B) Quantification of colony numbers formed by MDA‐MB‐231 under previously indicated conditions. Data are obtained from three independent experiments, with three technical replicates per condition. Statistical analyses were performed using a Mann–Whitney test. MDA = MDA‐MB‐231 cells. Scale bar = 600 μm. DMEM, Dulbecco'’s Modified Eagle Medium; hBP‐CM, hBP‐conditioned medium; TNBC, triple‐negative breast cancer.

Article Snippet: The human TNBC cell line MDA‐MB‐231 (female) was obtained from the American type culture collection (ATCC, HTB‐26, RRID:CVCL_0062, obtained in 2023).

Techniques: Control, MANN-WHITNEY, Modification

Journal: Oncology Letters

Article Title: Methylglyoxal-derived glycated albumin enhances the stemness potential of invasive ductal carcinoma-derived breast cancer stem-like cell line KAIMRC1

doi: 10.3892/ol.2026.15541

Figure Lengend Snippet: ALDH activity in triple-negative breast cancer MDA-MB-231 and invasive ductal carcinoma-derived KAIMRC1 cells measurements using a colorimetric assay. KAIMRC1 stem-like cells showed higher ALDH activity when compared with that of MDA-MB-231 cells. Data are presented as the mean ± SD from three independent experiments. Statistical significance relative to MDA-MB-231 cells is indicated as *P<0.05. ALDH, aldehyde dehydrogenase.

Article Snippet: The TNBC cell line MDA-MB-231 (cat. no. HTB-26, American Type Culture Collection) was maintained in complete DMEM supplemented with the aforementioned components.

Techniques: Activity Assay, Derivative Assay, Colorimetric Assay

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