Journal: Bioactive Materials

Article Title: Harnessing the gut–immune–joint axis: Oral microalgae-based thermoresponsive microspheres enhance intra-articular therapy for rheumatoid arthritis

doi: 10.1016/j.bioactmat.2026.01.037

Figure Lengend Snippet: Evaluation of the protective effects of CG@GelMA on FSN-induced intestinal epithelial barrier disruption in vitro . (A) Immunofluorescence staining of Claudin-1 in Caco-2 monolayers under different treatments. Scale bar: 100 μm. (B) Relative fluorescence intensity of Claudin-1. (C) Immunofluorescence staining of Occludin in Caco-2 monolayers. Scale bar: 100 μm. (D) Relative fluorescence intensity of Occludin. (E) Immunofluorescence staining of ZO-1 in Caco-2 monolayers. Scale bar: 100 μm. (F) Relative fluorescence intensity of ZO-1. (G) Schematic diagram of the Caco-2/RAW 264.7 Transwell co-culture system. (H) Relative TEER of Caco-2 cell monolayers after different treatments. (I) Relative fluorescence intensity of FD4 across Caco-2 monolayers under different treatments. Data are presented as means ± SD. Statistical significance: ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: RAW 264.7 macrophages (Procell, Wuhan, China), Caco-2 intestinal epithelial cells (Pricella, Wuhan, China), and IEC-6 small intestinal epithelial cells (ATCC, USA) were maintained in high-glucose DMEM supplemented with 10 % fetal bovine serum (AiTing, Hangzhou, China) and 1 % penicillin-streptomycin (Gibco, USA), with the medium for IEC-6 cells additionally containing 0.1 U/mL human insulin.

Techniques: Disruption, In Vitro, Immunofluorescence, Staining, Fluorescence, Co-Culture Assay

Journal: Journal of Cell Science

Article Title: OptoLoop – an optogenetic tool to probe the functional role of genome organization

doi: 10.1242/jcs.264574

Figure Lengend Snippet: Optogenetic manipulation of proximity between repetitive genomic loci. (A) Scheme of OptoLoop consisting of a fusion between dCas9 and the optogenetic protein CRY2. OptoLoop is targeted to specific genomic loci by introducing specific sgRNAs. CRY2–CRY2 interactions activated by blue light bridge targeted loci to form a chromatin loop. (B) Left panel, region of chromosome 19 showing sgIDR3 and sgTCF3 target sites, representative Hi-C contact map (data from ) and BACs used in DNA-FISH to label the IDR3 (magenta) and TCF3 loci (green). Right panel, mCherry channel images of U2OS dCas9–3XmCherry–CRY2 cells transfected with sgIDR3 and sgTCF3, kept in dark or illuminated with blue light for 3 h (1 s pulses every 10 s), and fixed. Scale bars: 5 µm. (C) Left panel, representative image of DNA-FISH for IDR3 and TCF3 with specific BAC FISH probes in U2OS cells. Right panel represents a single cell highlighted in left panel (yellow box); the expansion shows a single allele in this cell. Dashed line denotes the distance between the two FISH signals. Scale bars: 20 µm (left panel), 5 µm (right panel), 1 µm (expansion). (D) IDR3–TCF3 distances, calculated for U2OS dCas9–mCherry–CRY2 polyclonal cells transfected with indicated combinations of sgIDR3 and sgTCF3, kept under dark or illuminated for 3 h (1 s pulses every 10 s). Violin plot corresponds to a representative experiment, with black lines representing median distances. Bar plot represents means of two independent experiments. Each dot represents the median of typically 5000–10,000 alleles analyzed per experiment. (E) Fraction of alleles with IDR3-TCF3 distance <0.27 µm measured from DNA-FISH images for U2OS dCas9–mCherry–CRY2 polyclonal cells and three clones of U2OS dCas9–3XmCherry–CRY2 cells, transfected with indicated combinations of sgIDR3 and sgTCF3, and kept in dark or illuminated for 3 h (1 s pulses every 10 s). Each dot represents the fraction of typically 5000–10,000 alleles analyzed per experiment. Bars represent means of two or three independent experiments. (F) Measurement of cell-to-cell heterogeneity in loop formation. Bars with green shades: observed fraction of cells with none, one or both alleles with IDR3–TCF3 distance <0.27 µm obtained from a representative experiment shown in E with 2500–5000 cells analyzed per sample. Bars with magenta shades: expected fraction of cells with none, one or both alleles with IDR3–TCF3 distance <0.27 µm assuming that alleles from a same cell are independent between each other (Eqn 2). * P <0.05; *** P <0.001; ns, not significant [two-way ANOVAs followed by post-hoc Tukey tests (D,E); paired two-tailed t -test (E); chi-squared test (F)].

Article Snippet: NIH3T3 (mouse fibroblasts, ATCC #CRL-1658), U2OS (from human osteosarcoma, ATCC #HTB-96), HeLa (from human cervical adenocarcinoma, ATCC #CRM-CCL-2) and Lenti-X HEK-293T (from human embryonic kidney, cat. #632180 from Takara Bio, Japan) cell lines were cultured in Dulbecco's modified Eagle's medium (Gibco, Waltham, MA, USA) supplemented with 10% (15% for NIH3T3) fetal bovine serum (Gibco, Waltham, MA, USA) plus 100 IU/ml penicillin and 100 μg/ml streptomycin (Gibco, Waltham, MA, USA) at 37°C in a humidified atmosphere with 5% CO 2 .

Techniques: Hi-C, Transfection, Single Cell, Clone Assay, Two Tailed Test

Journal: Journal of Cell Science

Article Title: OptoLoop – an optogenetic tool to probe the functional role of genome organization

doi: 10.1242/jcs.264574

Figure Lengend Snippet: Benchmarking OptoLoop against a previous optogenetic manipulation tool. (A) Scheme of LADL consisting of soluble CRY2wt and a fusion between dCas9 and the CRY2 partner CIBN. dCas9–CIBN tethers specific genomic loci and light-activation induces both CRY2–CRY2 and CRY2–CIBN interactions bridging the targeted loci to form a loop. (B) Images of U2OS cells expressing dCas9–3XGFP–CIBN and mCherry fused to CRY2wt or CRY2olig, transfected with sgIDR3 and sgTCF3, and fixed after being kept under dark or illuminated with blue light pulses for 3 h (1 s pulses every 10 s). White arrows indicate FISH signals corresponding to IDR3–TCF3 loci. Scale bar: 5 µm. (C) Fraction of alleles with IDR3–TCF3 distance <0.27 µm measured from DNA-FISH images of U2OS cell lines stably expressing dCas9-3XGFP-CRY2 and mCherry-CRY2olig (LADL, clone #7) or dCas9-3XmCherry-CRY2 (OptoLoop, clone #3), transfected with sgIDR3 and sgTCF3, and kept under dark or illuminated with blue light for 3 h (1 s pulses every 10 s). Each dot represents the fraction of 5000–7000 alleles analyzed per experiment. Bars represent the means of three independent experiments. * P <0.05; ** P <0.01; *** P <0.001 (two-way ANOVA followed by post-hoc Tukey test).

Article Snippet: NIH3T3 (mouse fibroblasts, ATCC #CRL-1658), U2OS (from human osteosarcoma, ATCC #HTB-96), HeLa (from human cervical adenocarcinoma, ATCC #CRM-CCL-2) and Lenti-X HEK-293T (from human embryonic kidney, cat. #632180 from Takara Bio, Japan) cell lines were cultured in Dulbecco's modified Eagle's medium (Gibco, Waltham, MA, USA) supplemented with 10% (15% for NIH3T3) fetal bovine serum (Gibco, Waltham, MA, USA) plus 100 IU/ml penicillin and 100 μg/ml streptomycin (Gibco, Waltham, MA, USA) at 37°C in a humidified atmosphere with 5% CO 2 .

Techniques: Activation Assay, Expressing, Transfection, Stable Transfection

Journal: Tumour Virus Research

Article Title: HPV18E6 and CDK5 virus-host interaction is a prospective therapeutic target for HPV-positive cervical cancer

doi: 10.1016/j.tvr.2026.200339

Figure Lengend Snippet: CP681301 exerted a greater inhibition on the proliferation ability of HPV-positive than HPV-negative cancer cells. (A). A schematic diagram summarises the screen process of CDK5 inhibitors against HPV-positive cancer cell lines. (B-C). CP681301 against cell proliferation of five CSEC and HNSCC, HeLa, CaSki, SCC090, C33A, and SAS. Representative examples of the results from 3 independent experiments. (D). The growth curve indicates the cell proliferation of HeLa cells in the presence or the absence of CP681301. This proliferation of HeLa was assessed using CCK8 cell viability assay. Representative examples of the results from 3 independent experiments. (E). The growth curve indicates the cell proliferation of CaSki cells in the presence or the absence of CP681301. The proliferation of CaSki was assessed using CCK8 cell viability assay. Representative examples of the results from 3 independent experiments. (F). The growth curve indicates the cell proliferation of C33A cells in the presence or the absence of CP681301. The proliferation of CaSki was assessed using CCK8 cell viability assay. Representative examples of the results from 3 independent experiments.

Article Snippet: HeLa (HPV18 positive cervical cancer cells), CaSki (HPV16 positive cervical cancer cells), human embryonic kidney (HEK) 293 (HPV-null epithelial cells), C33A (HPV-null cervical cancer cells), U-2 OS (HPV-null human osteosarcorma cells), SAS (HPV-null human head and neck squamous cell carcinoma cell line), and SCC090 (HPV16-positive human HNSCC cell line) were purchased from the American Type Culture Collection (ATCC) and maintained in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS, GIBCO) at 37 °C in a humidified incubator with 5% CO 2 .

Techniques: Inhibition, Viability Assay

Journal: Tumour Virus Research

Article Title: HPV18E6 and CDK5 virus-host interaction is a prospective therapeutic target for HPV-positive cervical cancer

doi: 10.1016/j.tvr.2026.200339

Figure Lengend Snippet: Inhibition of CDK5 dampens cancer phenotypes of HPV-positive cervical cancer cells. (A) (i). Representative images compare the cell migration of HeLa after scratching. Cells were seeded into 6-well plates and were scratched when they reached 100% confluence, then incubated with CP681301 (0.6, 1.5 μM) or DMSO for 24 h. Cell migration was observed and recorded through time-lapse microscopy at different time points (0, 6, 12, 24 h). (ii) The bar charts compare the relative coverage of the scratched area of HeLa upon treatment with DMSO or CP681301 (n = 3). (B) (i) The representative images show the whole-well view and colony morphology of the colonies formed by HeLa cells. Cells were seeded into 0.35% agarose containing DMEM with 20% FBS and were grown for 14 days. The agarose was stained with crystal violet. The colony morphology was observed and recorded under an optical microscope at × 100 magnification. (ii) The bar charts compare the colony number of HeLa cells upon treatment with CP681301 or DMSO (n = 3). (C) (i) The representative images compare the cell invasion of HeLa cells upon treatment with CP681301 or DMSO. Cells were seeded into the invasion chamber with serum-free medium. Cell invasion was observed and recorded through an optical microscope. (ii) The bar graphs compare the relative cell invasion abilities of HeLa cells upon treatment with CP681301 and DMSO (n = 3). Error bars indicate SEM. ∗p < 0.05; ∗∗p < 0.01. ∗∗∗p < 0.001.(D) (i) The representative images compare the cell invasion of C33A cells upon treatment with CP681301 or DMSO. Cells were seeded into the invasion chamber with serum-free medium. Cell invasion was observed and recorded through an optical microscope. (ii) The bar graphs compare the relative cell invasion abilities of C33A cells upon treatment with CP681301 and DMSO (n = 3). Error bars indicate SEM. ∗p < 0.05; ∗∗p < 0.01. ∗∗∗p < 0.001.

Article Snippet: HeLa (HPV18 positive cervical cancer cells), CaSki (HPV16 positive cervical cancer cells), human embryonic kidney (HEK) 293 (HPV-null epithelial cells), C33A (HPV-null cervical cancer cells), U-2 OS (HPV-null human osteosarcorma cells), SAS (HPV-null human head and neck squamous cell carcinoma cell line), and SCC090 (HPV16-positive human HNSCC cell line) were purchased from the American Type Culture Collection (ATCC) and maintained in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS, GIBCO) at 37 °C in a humidified incubator with 5% CO 2 .

Techniques: Inhibition, Migration, Incubation, Time-lapse Microscopy, Staining, Microscopy

Journal: Tumour Virus Research

Article Title: HPV18E6 and CDK5 virus-host interaction is a prospective therapeutic target for HPV-positive cervical cancer

doi: 10.1016/j.tvr.2026.200339

Figure Lengend Snippet: CP681301 treatment destabilized E6, reduced E6AP and rescued p53. (A). HeLa and C33A cell lines were treated with CP681301 at the concentrations of 0.6 μM and 1.5 μM for 24 h. The protein extracts were subjected to Western blotting for the detection of CDK5, 18E6, E6AP, p53, and β-actin proteins. (B). CP681301 disrupted the 18E6-E6AP protein complex. The inhibition of CP681301 was detected using the GST-pull down assay, where CP681301 was incubated with the purified GST-18E6 protein and Flag-E6AP protein for 2h. After extensive washing, the bound E6AP protein was detected via Western blotting using an anti-Flag antibody. The immunoblot (IB) on the upper panel shows the interaction of E6AP with GST-18E6, while the lower panel shows the Ponceau S stained of the blot.

Article Snippet: HeLa (HPV18 positive cervical cancer cells), CaSki (HPV16 positive cervical cancer cells), human embryonic kidney (HEK) 293 (HPV-null epithelial cells), C33A (HPV-null cervical cancer cells), U-2 OS (HPV-null human osteosarcorma cells), SAS (HPV-null human head and neck squamous cell carcinoma cell line), and SCC090 (HPV16-positive human HNSCC cell line) were purchased from the American Type Culture Collection (ATCC) and maintained in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS, GIBCO) at 37 °C in a humidified incubator with 5% CO 2 .

Techniques: Western Blot, Inhibition, Pull Down Assay, Incubation, Purification, Staining

Journal: Tumour Virus Research

Article Title: HPV18E6 and CDK5 virus-host interaction is a prospective therapeutic target for HPV-positive cervical cancer

doi: 10.1016/j.tvr.2026.200339

Figure Lengend Snippet: CDK5 forms a complex with E6 predominantly in the nucleus. (A) (i). The represented immunoblot image of CDK5 protein binding with 16E6/E7 and 18E6/E7 proteins. GST pull-down assay was performed by incubating the indicated purified GST fusion proteins with CDK5 protein. After extensive washing, the bound CDK5 protein was detected via Western blotting using an anti-CDK5 antibody. The immunoblot (IB) on the upper panel shows the interaction of CDK5 with GST-16 E6/E7 and GST-18 E6/E7, while the lower panel shows the Ponceau S stain of the blot. (ii) The bar graph shows the quantification of the relative level of CDK5 to GST empty protein indicated from 3 independent experiments (n = 3). Quantitation was performed using ImageJ software, and the statistical analysis was performed using Graphpad Prism 8. (B). Representatives immunoblot of Co-immunoprecipitation shows that CDK5 binds with 16E6 and 18E6. The HA-16E6, HA-18E6, and His-CDK5 were transfected into the HEK293 cells. After 24 h, the lysates from cells were analyzed by western blotting using anti-CDK5, anti-HA specific antibodies. Data were expressed as mean ± standard error of the mean (SEM, ∗: p < 0.05, ∗∗: p < 0.01, ∗∗∗: p < 0.005, ∗∗∗∗: p < 0.0001). (C). U-2 OS cells were transfected with pcDNA3.1: His-CDK5 (His-CDK5) and pcDNA3.1: HA-16E6 (HA-16E6) and HA-18E6 (HA-18E6) plasmids. The cells were fixed and incubated with primary antibodies (anti-His, anti-HA), followed by incubation with the relevant Alexa Fluor 568-conjugated anti-rabbit and Alexa Fluor 488-conjugated anti-mouse secondary antibodies. The cells were then counterstained with 4,6-diamidino-2-phenylindole (DAPI). The Z-stacking images for subcellular expression of CDK5 (Red) and E6 (Green) were examined using the Nikon fluorescence microscope. Cellular localization of CDK5 and E6 was visualized by a fluorescent microscope under 1000× magnification.

Article Snippet: HeLa (HPV18 positive cervical cancer cells), CaSki (HPV16 positive cervical cancer cells), human embryonic kidney (HEK) 293 (HPV-null epithelial cells), C33A (HPV-null cervical cancer cells), U-2 OS (HPV-null human osteosarcorma cells), SAS (HPV-null human head and neck squamous cell carcinoma cell line), and SCC090 (HPV16-positive human HNSCC cell line) were purchased from the American Type Culture Collection (ATCC) and maintained in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS, GIBCO) at 37 °C in a humidified incubator with 5% CO 2 .

Techniques: Western Blot, Protein Binding, Pull Down Assay, Purification, Staining, Quantitation Assay, Software, Immunoprecipitation, Transfection, Incubation, Expressing, Fluorescence, Microscopy

Journal: Bioactive Materials

Article Title: Tailoring nanotopography and antibacterial properties of calcium phosphate bone grafts via fluoride incorporation

doi: 10.1016/j.bioactmat.2025.12.026

Figure Lengend Snippet: (a) Cell proliferation of SaOS-2 cells onto the different CaP discs for 4 h, 7, 14 and 21 days. (b) ALP activity of SaOS-2 cells cultured onto the CaP substrates for 4 h, 7, 14 and 21 days. The same letter indicates no statistically significant differences for the same group at different time points while the same number denotes no statistically significant differences for each time point among all samples. (p < 0.05).

Article Snippet: SaOS-2 cell line (HTB-85), RAW 264.7 cell line (TIB-71), Pseudomonas aeruginosa (ATCC-27853) and Staphylococcus aureus (ATCC-25923) were purchased from American Type Culture Collection (VA, USA).

Techniques: Activity Assay, Cell Culture

Journal: Bioactive Materials

Article Title: Tailoring nanotopography and antibacterial properties of calcium phosphate bone grafts via fluoride incorporation

doi: 10.1016/j.bioactmat.2025.12.026

Figure Lengend Snippet: Merged CLSM images of SaOS-2 cells cultured for 4 h and 7 days on the nanostructured CaP substrates, as well as the Flat and Ti controls. Actin filaments were stained with Alexa Fluor™ 546 phalloidin (orange fluorescence signal) and nuclei with DAPI (blue fluorescence signal).

Article Snippet: SaOS-2 cell line (HTB-85), RAW 264.7 cell line (TIB-71), Pseudomonas aeruginosa (ATCC-27853) and Staphylococcus aureus (ATCC-25923) were purchased from American Type Culture Collection (VA, USA).

Techniques: Cell Culture, Staining, Fluorescence

Journal: Bioactive Materials

Article Title: Tailoring nanotopography and antibacterial properties of calcium phosphate bone grafts via fluoride incorporation

doi: 10.1016/j.bioactmat.2025.12.026

Figure Lengend Snippet: (a) mRNA expression of osteogenic genes ALPL, RUNX2 and SPP1 of SaOS-2 cells cultured directly onto the CaP substrates for 1, 3, 7 and 14 days, determined by real-time PCR (n = 3). (b) Interaction of RAW 246.7 cells with treated discs up to 7 days in cell culture. i) mRNA expression of pro-inflammatory genes TNF, IL1B and IL6 of cells in the direct cell culture, determined by real-time PCR for 1, 3 and 7 days (n = 3). All values are relativized to values of cells at day 1. ii) Protein expression level after 7 days of direct cell culture on the treated discs, measured by inflammation antibody array. Values of protein signal are quantified by image analysis and relativized to control. In Fig. a) and bi), the same letter indicates no statistically significant differences for the same group at different time points while the same number denotes no statistically significant differences for each time point among all samples. (p < 0.05).

Article Snippet: SaOS-2 cell line (HTB-85), RAW 264.7 cell line (TIB-71), Pseudomonas aeruginosa (ATCC-27853) and Staphylococcus aureus (ATCC-25923) were purchased from American Type Culture Collection (VA, USA).

Techniques: Expressing, Cell Culture, Real-time Polymerase Chain Reaction, Ab Array, Control

Journal: Bioactive Materials

Article Title: Tailoring nanotopography and antibacterial properties of calcium phosphate bone grafts via fluoride incorporation

doi: 10.1016/j.bioactmat.2025.12.026

Figure Lengend Snippet: Co-culture of P. aeruginosa and SaOS-2 cells on the nanostructured CaP discs, as well as Flat and Ti controls. a) Merged CLSM images of: i) a pre-implantation infection model, where the samples were first incubated for 6 h with P. aeruginosa and subsequently SaOS-2 cells were cultured for 24 h; or ii) post-implantation infection model, where SaOS-2 cells were first cultured for 24 h on the substrates, which were subsequently incubated for 6 h with P. aeruginosa . Actin filaments were stained with Alexa Fluor™ 546 phalloidin (orange fluorescence signal) and the nuclei with DAPI (blue fluorescence signal). (b) Orthogonal CLSM images showing simultaneous co-visualization of cells and bacteria stained with Alexa Fluor™ 546 phalloidin (orange fluorescence signal), the nuclei with DAPI (blue fluorescence signal) and SYTO-9 (green fluorescence signal). (c) Dead percentage of P. aeruginosa onto the different CaP substrates and the controls, for both the pre-implantation infection i) and the post-implantation ii) infection models. n = 3; ns and ∗ indicate significance at p > 0.05 and p ≤ 0.05, respectively.

Article Snippet: SaOS-2 cell line (HTB-85), RAW 264.7 cell line (TIB-71), Pseudomonas aeruginosa (ATCC-27853) and Staphylococcus aureus (ATCC-25923) were purchased from American Type Culture Collection (VA, USA).

Techniques: Co-Culture Assay, Infection, Incubation, Cell Culture, Staining, Fluorescence, Bacteria

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: 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