mouse k7m2 wt cells  (Procell Inc)

Journal: Advanced Science

Article Title: Osteosarcoma Cell‐Derived Migrasomes Promote Macrophage M2 Polarization to Aggravate Osteosarcoma Proliferation and Metastasis

doi: 10.1002/advs.202409870

Figure Lengend Snippet: MANPs promote macrophage phagocytosis and M2 polarization via Milk fat globule‐EGF factor 8 (MFGE8). A‐H) 4D proteome sequencing analysis of cell body, migrasome and MANP. (A‐D) Heatmaps and volcano plots of DEGs in protein‑seq analysis of cell body, migrasome and MANP ( n = 3). (E, F) Heatmaps of TSPAN proteins, integrin proteins, and histones in cell body, migrasome and MANP ( n = 3). (G) Venn diagram showing proteins co‐enriched by migrasomes and MANPs. (H) Relative expression of MFGE8 in cell body, migrasome and MANP ( n = 3). I) Immunostaining images of WGA (green) and MFGE8 (red) in OS cells ( n = 3). Magnified images of migrasomes and MANPs are shown on the right. Low magnification: scale bars = 10 µm; high magnification: scale bars = 1 µm. J,K) qRT‐PCR and WB analysis of MFGE8 knockdown efficiency in K7M2 wt cells ( n = 3). L,M) WB analysis of MFGE8 knockdown efficiency in migrasomes and MANPs ( n = 3). N‐Q) BMDMs treated with 10 µg mL −1 sh‐NC MANPs or sh‐MFGE8 MANPs for 24 h. (N, O) Representative immunostaining images and quantitative analysis of BMDMs (red) phagocytosis of microbeads (green) ( n = 3). Low magnification: scale bars = 100 µm; high magnification: scale bars = 50 µm. (P, Q) Flow cytometry analysis of the proportion of BMDMs (APC) that phagocytose apoptotic OS cells (CFSE) and quantitative analysis ( n = 3). R‐T) BMDMs treated with 10 µg mL −1 sh‐NC MANPs or sh‐MFGE8 MANPs for 24 h, followed by co‐incubation with apoptotic OS cells for 24 h. (R) qRT‐PCR analysis of the expression of M1 polarization and M2 polarization markers in BMDMs ( n = 3). (S, T) Flow cytometry analysis of the proportion of M2‐type macrophages and quantitative analysis ( n = 3). Results were shown as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001. One‐way ANOVA test or two‐way ANOVA test was used for multivariate analysis. Unpaired t‐tests were used for the comparison of two groups.

Article Snippet: The human OS cell lines 143b and MG63 and the mouse OS cell line K7M2 wt were obtained from the American Type Culture Collection (ATCC), and the OS cell lines were cultured in Dulbecco's modified Eagle's medium (DMEM, C11995500BT, Gibco, USA) supplemented with 1% penicillin/streptomycin (P/S, 15140122, Gibco, USA) and 10% fetal bovine serum (FBS, 10091148, Gibco, USA).

Techniques: Sequencing, Expressing, Immunostaining, Quantitative RT-PCR, Knockdown, Flow Cytometry, Incubation, Comparison

Journal: Molecular Therapy Oncology

Article Title: Trabectedin promotes oncolytic virus antitumor efficacy, viral gene expression, and immune effector function in models of bone sarcoma

doi: 10.1016/j.omton.2024.200886

Figure Lengend Snippet: Trabectedin improves oHSV antitumor efficacy with reproducibility in immunocompetent osteosarcoma mouse models The best response for each treated tumor through 28 days, the average tumor burden, spider plots, and individual body weights are shown for treated osteosarcoma models (A) K7M2 and (B) F420 in immunocompetent BALB/c and B6-albino mice, respectively. Body weight plots include mice that were excluded from the tumor burden analyses due to early non-tumor-related endpoints. PBS and oHSV (1.0 × 10 8 pfu) were given i.Tu. on days 0, 2, and 4. Trabectedin (0.15 mg/kg) was given i.v. on days 0 and 7. Statistical analyses of the disease control rates (CR + PR + SD) were performed using a pairwise Fisher’s exact test with p values adjusted using the Benjamini-Hochberg procedure; ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001. Summarized data with error bars depict mean ± SEM.

Article Snippet: The A673 human Ewing sarcoma cell line (Cat# CCL-81), Vero green monkey kidney cell line (Cat# CRL-1598), and K7M2 mouse osteosarcoma cell line (Cat# CRL-2836) were purchased from the American Type Culture Collection (ATCC) (Manassas, VA).

Techniques: Control

Journal: Molecular Therapy Oncology

Article Title: Trabectedin promotes oncolytic virus antitumor efficacy, viral gene expression, and immune effector function in models of bone sarcoma

doi: 10.1016/j.omton.2024.200886

Figure Lengend Snippet: Trabectedin reduces immune suppression and enhances effector cell activation to synergize with oHSV (A) scRNA-seq UMAP plot depicts the treated K7M2 tumor microenvironments. Datasets from all treatment groups were merged with no need for batch correction. (B) Bar plot displays the normalized enrichment score (NES) of the significant KEGG pathways from gene set enrichment analysis in NK and CD8 T cells from the tumor microenvironment. (C) FACS of tumor-infiltrating immune cells showed decreased immunosuppressive cells and increased activated T cells post-treatment. The p values calculated via one-way ANOVA with Dunnett’s multiple comparisons test are shown (wherein each treatment cohort was compared with the combination cohort). Error bars depict mean ± SEM. (D) Violin plots show increased cytotoxic gene expression in all NK and T cells of combination-treated K7M2 (arrow indicates control for comparison, percent change = 100∗(2 log2FC -1)). All treatment comparisons with a percent change value over 20% were statistically significant with p -adjusted ≤ 0.01. PBS and oHSV (1.0 × 10 8 pfu) were given i.Tu. on days 0 and 2. Trabectedin (0.15 mg/kg) was given i.v. on day 0. scRNA-seq samples were collected on day 3. Flow cytometry samples were collected on day 7.

Article Snippet: The A673 human Ewing sarcoma cell line (Cat# CCL-81), Vero green monkey kidney cell line (Cat# CRL-1598), and K7M2 mouse osteosarcoma cell line (Cat# CRL-2836) were purchased from the American Type Culture Collection (ATCC) (Manassas, VA).

Techniques: Activation Assay, Gene Expression, Control, Comparison, Flow Cytometry

Journal: Molecular Therapy Oncology

Article Title: Trabectedin promotes oncolytic virus antitumor efficacy, viral gene expression, and immune effector function in models of bone sarcoma

doi: 10.1016/j.omton.2024.200886

Figure Lengend Snippet: Combination efficacy in immunocompetent osteosarcoma models results from augmentation of antitumor NK and T cell responses The best response for each treated tumor through 28 days, the average tumor burden, and spider plots tracking individual tumor volumes over the full study period are displayed for (A) K7M2 tumor-bearing nude mice (lack T and B cells), (B) K7M2-bearing BALB/c mice treated with antibody-mediated CD4 and CD8 T cell depletion (given i.v. on days 0, 4, 8, 12, 16, 20, 24, and 28), (C) K7M2-bearing BALB/c mice treated with anti-asialo NK cell depletion (given i.v. on days 0, 7, 14, and 21). PBS and oHSV (1.0 × 10 8 pfu) were given i.Tu. on days 0, 2, and 4. Trabectedin (0.15 mg/kg) was given i.v. on days 0 and 7. Statistical analyses of the disease control rates (CR + PR + SD) were performed using a pairwise Fisher’s exact test with p values adjusted using the Benjamini-Hochberg procedure; ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001. Summarized data with error bars depict mean ± SEM.

Article Snippet: The A673 human Ewing sarcoma cell line (Cat# CCL-81), Vero green monkey kidney cell line (Cat# CRL-1598), and K7M2 mouse osteosarcoma cell line (Cat# CRL-2836) were purchased from the American Type Culture Collection (ATCC) (Manassas, VA).

Techniques: Control