Journal: Journal of Extracellular Vesicles

Article Title: Perivascular cell‐derived extracellular vesicles stimulate colorectal cancer revascularization after withdrawal of antiangiogenic drugs

doi: 10.1002/jev2.12096

Figure Lengend Snippet: Reduced tumor revascularization after ablation of NG2 + perivascular cells. (a) Therapeutic schedule for treatment with vehicle and regorafenib. Subcutaneous inoculation of Colon26 cancer cells in NG2‐tk mice and WT littermates, with daily GCV (50 mg/kg) or saline injections when tumors reached approximately 200 mm 3 . Tumor vasculatures were analyzed at days 7 (Day 7) and 12 (withdrawal 5 d) after the indicated treatments. Withdrawal 5 d, 5 days after treatment withdrawal. (b) Representative images of CD31 + (red) endothelial cell, NG2 + (green) perivascular cell immunostaining, and DAPI (blue) for nuclear staining. Scale bars, 50 μm. (c) Quantification of microvessel density (CD31 density) and perivascular cell number (NG2 density) in Colon26 tumors (n = 6). Data are present as mean ± SEM. * P < 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: Human colorectal cancer HT‐29 and HCT116 cells (ATCC), and mouse colorectal cancer Colon26 cells (JCRB Cell Bank) were cultured in DMEM (Invitrogen) supplemented with 10% FBS and 1% penicillin‐streptomycin (HyClone).

Techniques: Saline, Immunostaining, Staining

Journal: Journal of Extracellular Vesicles

Article Title: Perivascular cell‐derived extracellular vesicles stimulate colorectal cancer revascularization after withdrawal of antiangiogenic drugs

doi: 10.1002/jev2.12096

Figure Lengend Snippet: Tumor perivascular cells regulate EPC recruitment into tumors during the revascularization phase. (a and b) Quantification of the proportional changes of recruited EPCs in HT‐29 tumors (n = 6). Withdrawal 3/7 d, 3/7 days after AA‐TKI withdrawal. W‐3/7 d, 3/7 days after AA‐TKI withdrawal. (c) Quantification of the proportional changes of recruited EPCs in Colon26 tumors from NG2‐tk+GCV and WT+GCV mice after the indicated treatments (n = 6). (d) Pearson correlation analysis of EPC proportion and CD31 density in (d, left) HT‐29 tumors after treatment with AA‐TKIs for 7 days or withdrawal for 7 days; or in (d, right) Colon26 tumors from WT+GCV mice after treatment with regorafenib for 7 days or withdrawal for 5 days. (e) Pearson correlation analysis of (e, left) EPC proportion and perivascular cell number (α‐SMA density) in HT‐29 tumors after treatment with AA‐TKIs for 7 days or withdrawal for 7 days, or (e, right) EPC proportion and perivascular cell number (NG2 density) in Colon26 tumors from WT+GCV and NG2‐tk+GCV mice after treatment with regorafenib for 7 days or withdrawal for 5 days. (f) Representative images of PKH67‐labeled EPCs (green) recruited into HT‐29 tumors (mice 1) and Colon26 tumors (mice 2). DAPI (blue) was used for nuclear staining. (g) HT‐29 tumor‐bearing mice were treated with regorafenib + anti‐PDGFRβ for 7 days or (H) Colon26 tumor‐bearing NG2‐tk+GCV mice were treated with regorafenib for 7 days, followed by tail vein injection of 5 × 10 5 EPCs every other day. Tumor microvessel density was determined after 5 days. Representative images of CD31 + (red) endothelial cell, PDGFRβ + or NG2 + (green) perivascular cell immunostaining, and DAPI (blue) for nuclear staining. Quantification of CD31 density (n = 6). Data are present as mean ± SEM. Scale bars, 50 μm. * P < 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: Human colorectal cancer HT‐29 and HCT116 cells (ATCC), and mouse colorectal cancer Colon26 cells (JCRB Cell Bank) were cultured in DMEM (Invitrogen) supplemented with 10% FBS and 1% penicillin‐streptomycin (HyClone).

Techniques: Labeling, Staining, Injection, Immunostaining

Journal: Journal of Extracellular Vesicles

Article Title: Perivascular cell‐derived extracellular vesicles stimulate colorectal cancer revascularization after withdrawal of antiangiogenic drugs

doi: 10.1002/jev2.12096

Figure Lengend Snippet: TPC‐EVs promote EPC recruitment into tumors to facilitate tumor revascularization. (a‐c) The characterization and quantification of TPC‐EVs by (a) TEM, (b) particle size, and (c) Western blotting analyses. (d) USPIO‐loaded TPC‐EVs (100 μg) were intravenously injected into mice and TPC‐EV incorporation into bone marrow was detected by magnetic resonance imaging analysis. White arrows indicated the accumulation of USPIO‐loaded TPC‐EVs. USPIO, ultrasmall superparamagnetic iron oxide nanoparticles. (e) PKH26‐labeled TPC‐EVs (100 μg) were intravenously injected into mice. Barrow cells were harvested after 24 h and then fixed and incubated with red blood cell lysis buffer. Representative images of incorporated TPC‐EVs (red) in marrow cells. Scale bar, 20 μm. (f) HT‐29 tumor‐bearing mice received the treatment of regorafenib or axitinib for 7 days, followed by tail vein injection with PBS (Vehicle) or TPC‐EVs (100 μg) for 4 days. Then, EPC recruitment and tumor vasculature were analyzed. (g) Quantification of the proportional changes of recruited EPCs in HT‐29 tumors (n = 6). (h) Representative images of CD31 + (red) endothelial cell, α‐SMA + (green) perivascular cell immunostaining, and DAPI (blue) for nuclear staining. Scale bar, 50 μm. (i) Quantification of microvessel density (CD31 density) and perivascular cell number (α‐SMA density) in HT‐29 tumors (n = 6). Colon26 tumor‐bearing NG2‐tk+GCV mice were treated with regorafenib for 7 days, followed by tail vein injection of Vehicle or TPC‐EVs (100 μg) for 4 days. (j) Quantification of the proportional changes of recruited EPCs in Colon26 tumors (n = 6). (k) Representative images of CD31 + (red) endothelial cell, NG2 + (green) perivascular cell immunostaining, and DAPI (blue) for nuclear staining. Scale bar, 50 μm. (l) Quantification of microvessel density (CD31 density) and perivascular cell number (NG2 density) in Colon26 tumors (n = 6). Data are present as mean ± SEM. * P < 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: Human colorectal cancer HT‐29 and HCT116 cells (ATCC), and mouse colorectal cancer Colon26 cells (JCRB Cell Bank) were cultured in DMEM (Invitrogen) supplemented with 10% FBS and 1% penicillin‐streptomycin (HyClone).

Techniques: Western Blot, Injection, Magnetic Resonance Imaging, Labeling, Incubation, Red Blood Cell Lysis, Immunostaining, Staining

Journal: Nucleic Acids Research

Article Title: Separate transcription and splicing gene networks are linked and coordinated by the pRb–E2F pathway

doi: 10.1093/nar/gkag016

Figure Lengend Snippet: PRMT5 regulates pRb–E2F transcriptional and splicing networks in vivo . ( A ) Differential changes in splicing between Colon26 tumours isolated from mice treated with T1-44, or treated with DMSO, displayed as a heatmap of PSI values (Ψ, PSI) for all significant differential splicing events (FDR < 0.05). Each column of the heatmap represents PSI values of one splice event across all samples. Data clustering used the Canberra distance method. These data were generated from four mice per treatment group. ( B ) The bar chart displays the breakdown of statistically significant (FDR < 0.05) differential splicing events observed in Colon26 tumours from mice treated with T1-44 as compared to tumours from mice treated with DMSO. SE, skipped/cassette exon; RI, retained intron; MXE, mutually exclusive exons; A5SS, alternative 5′ splice site; A3SS, alternative 3′ splice. These data were derived from the analysis in Fig. . A volcano plot displaying values of log2 fold change (log2FC) and –log10 P -values for DEGs identified between Colon26 tumours from mice treated with T1-44 as compared to tumours from mice treated with DMSO. Red colour represents genes upregulated by T1-44 treatment, whilst blue colour represents genes downregulated by T1-44 treatment. Grey colour represents genes that fell below the fold change or statistical cut-off applied (padj < 0.05, log2FC > 0.58). These data were derived from the same RNA-seq analysis used to generate Fig. . ( D ) A venn diagram displaying the overlap of total genes from the Colon26 tumour RNA-seq that score either as significantly differentially expressed (DEGs: padj < 0.05, log2FC > 0.58), significantly differentially spliced (AS: FDR < 0.05), or fall into both categories. These data were derived from the analysis in Fig. and . ( E ) RNA extracted from Colon26 tumours of mice treated with T1-44 or DMSO was used in an RT-PCR experiment to measure the inclusion of the indicated exons in RNA transcripts. Displayed is the mean inclusion/exclusion ratio for each gene, with individual data points displayed. Significance was calculated by Student’s t -test ( n = 4 mice from each treatment group).

Article Snippet: Mouse Colon26 cells were used by Charles Rivers Laboratories (Wilmington, USA) for out-sourced tumour challenge models [ ].

Techniques: In Vivo, Isolation, Generated, Derivative Assay, RNA Sequencing, Reverse Transcription Polymerase Chain Reaction

Journal: Molecular Oncology

Article Title: Immune modulation underpins the anti‐cancer activity of HDAC inhibitors

doi: 10.1002/1878-0261.12953

Figure Lengend Snippet: Genome‐wide analysis on CXD101‐treated colon26 cells. (A) Heat map of differential gene expression observed in colon26 cells. The heat map shows 2514 significantly DEGs between CXD101‐treated (2.7 µ m for 72 h) and control (DMSO) colon26 cells. Normalised rlog‐transformed gene expression values corresponding to significantly expressed genes (FDR < 0.01 and |log2(FC)| > 1 were mean‐centred by rows. Each row of the heat map represents transformed expression values of one DEG across all samples (blue, low expression; red, high expression). Genes associated with immune system‐related KEGG pathways (see panel (B)) are indicated in violet on a separate panel (in.mm kegg) on the left of the heat map (see also Dataset ); n = 3. (B) Significantly over‐represented KEGG concepts encompassing pathways associated with significant differences in expression change upon CXD101 treatment in colon26 cells. Heat maps show PGSEA statistic ( Z ‐score), which characterises how much the mean of the fold changes for genes in a certain pathway deviates from the mean observed in all the genes between CXD101 treatment and the control (DMSO) groups. Blue indicates gene sets with decreased expression, while red corresponds to those with increased; n = 3. (C) Significantly over‐represented GO terms encompassing biological processes or cellular components associated with significant differences in expression change upon CXD101 treatment in colon26 cells. Heat maps show PGSEA statistic ( Z ‐score), which characterises how much the mean of the fold changes for genes in a certain pathway deviates from the mean observed in all the genes between CXD101 treatment and the control (DMSO) groups. Blue indicates gene sets with decreased expression, while red corresponds to those with increased; n = 3. (D) Gene expression heat map showing significantly DEGs associated with ‘Antigen processing and presentation’ KEGG pathway, referred to as AP signature. Gene expression values were transformed using approach outlined above (see panel A). (E) Heat map of differential expression showing significantly DEGs associated with ‘Natural Killer Cell‐Mediated Cytotoxicity’ KEGG pathway, referred to as NK signature. Gene expression values were transformed using approach outlined above (see panel A). (F) qRT‐PCR of genes identified in panels D and E (i and ii, respectively) in colon26 cells treated for 3 days with 2.7 µ m CXD101 or DMSO control (Student's t ‐test; * P < 0.05, error bars indicate SD); (iii) an immunoblot of colon26 cells is included to demonstrate input protein levels for H3acK9; actin included as a loading control; n = 3.

Article Snippet: Human colorectal adenocarcinoma SW620 (ATCC ® CCL‐227; RRID:CVCL_0547) and HCT116 (ATCC ® CCL‐247; RRID:CVCL_0291), human breast cancer MCF7 (ATCC ® HTB‐22; RRID:CVCL_0031), human lung cancer A549 (ATCC ® CCL‐185; RRID:CVCL_JK07) and the mouse colon carcinoma cell line colon26 (ATCC ® CRL‐2639; RRID:CVCL_7255) were obtained from ATCC (Manassas, VA, USA).

Techniques: Genome Wide, Gene Expression, Control, Transformation Assay, Expressing, Quantitative Proteomics, Quantitative RT-PCR, Western Blot

Journal: Molecular Oncology

Article Title: Immune modulation underpins the anti‐cancer activity of HDAC inhibitors

doi: 10.1002/1878-0261.12953

Figure Lengend Snippet: Genome‐wide analysis on CXD101‐treated colon26 tumours. (A) Schematic representation of the experiment with CXD101 in colon26 tumours (i). Balb/c mice were treated with orally administrated CXD101 at 50 mg·kg −1 for 14 days with respect to vehicle‐only control; n = 10 per group; (ii) relative tumour growth volume in CXD101‐treated and nontreated Balb/c mice presented as a mean value (Student's t ‐test of the values at day 14; * P < 0.05); (iii) scatter plots of relative tumour volume of individual mouse at day 14 ( t ‐test; * P < 0.05); (iv) relative body weight representation of CXD101‐treated and nontreated Balb/c mice presented as a mean value. (B) Heat map of differential expression. The heat map shows 1036 significantly DEGs between CXD101‐treated ( n = 4) and control ( n = 3) colon26 syngeneic tumour samples. Normalised rlog‐transformed gene expression values corresponding to significantly expressed genes (FDR < 0.01 and |log2(FC)| > 1 were mean‐centred by rows). Each row of the heat map represents transformed expression values of one DEG across all samples (blue, low expression; red, high expression). Genes associated with immune system‐related KEGG pathways (see panel (C)) are indicated in violet on a separate panel (in.mm kegg) on the left of the heat map (see also Dataset ). (C) Significantly over‐represented KEGG concepts encompassing pathways associated with significant differences in expression change upon CXD101 treatment of the tumours. Heat maps show PGSEA statistic ( Z ‐score), which characterises how much the mean of the fold changes for genes in a certain pathway deviates from the mean observed in all the genes between CXD101 treatment ( n = 4) and the control (DMSO, n = 3) groups. Blue indicates gene sets with decreased expression, while red corresponds to those which were increased. (D) Significantly over‐represented GO terms encompassing biological processes or cellular components associated with significant differences in expression change upon CXD101 treatment of the tumours. Heat maps show PGSEA statistic ( Z ‐score), which characterises how much the mean of the fold changes for genes in a certain pathway deviates from the mean observed in all the genes between CXD101 treatment ( n = 4) and the control (DMSO, n = 3) groups. Blue indicates gene sets with decreased expression, while red corresponds to those with increased. (E) Gene expression heat map showing significantly DEGs associated with ‘Antigen processing and presentation’ KEGG pathway, the AP signature. Gene expression values were transformed using approach outlined above (see panel B). (F) Heat map of differential expression showing significantly DEGs associated with ‘Natural Killer Cell‐Mediated Cytotoxicity’ KEGG pathway, the NK signature. Gene expression values were transformed using approach outlined above (see panel B). (G) qRT‐PCR validation of genes identified in panels E and F (i and ii, respectively) in colon26 syngeneic tumour RNA treated for 14 days with 50 mg·kg −1 CXD101 or DMSO control; n = 3 (Student's t ‐test; * P < 0.05, error bars indicate SD).

Article Snippet: Human colorectal adenocarcinoma SW620 (ATCC ® CCL‐227; RRID:CVCL_0547) and HCT116 (ATCC ® CCL‐247; RRID:CVCL_0291), human breast cancer MCF7 (ATCC ® HTB‐22; RRID:CVCL_0031), human lung cancer A549 (ATCC ® CCL‐185; RRID:CVCL_JK07) and the mouse colon carcinoma cell line colon26 (ATCC ® CRL‐2639; RRID:CVCL_7255) were obtained from ATCC (Manassas, VA, USA).

Techniques: Genome Wide, Control, Quantitative Proteomics, Transformation Assay, Gene Expression, Expressing, Quantitative RT-PCR, Biomarker Discovery

Journal: Molecular Oncology

Article Title: Immune modulation underpins the anti‐cancer activity of HDAC inhibitors

doi: 10.1002/1878-0261.12953

Figure Lengend Snippet: CXD101 affects the TME of colon26 tumours. (A) Representative examples of immunohistochemical staining of H3AcK9 in colon26 tumours collected from Balb/c mice at 14 days treated with 50 mg·kg −1 CXD101 and nontreated control (see experiment in Fig. ). Original magnification: 20×, scale bar, 50 μm; and 63×; scale bar, 16 μm. n = 4. (B) As above, but immunohistochemical staining was performed with anti‐CD4. (C) As above, but immunohistochemical staining was performed with anti‐CD8. (D) As above, but immunohistochemical staining was performed with anti‐FoxP3. (E) As above, but immunohistochemical staining was performed with anti‐CD68 gene. (F) As above, but immunohistochemical staining was performed with anti‐CD163. (G) As above, but immunohistochemical staining was performed with anti‐NKp46. (H) Results were quantified by imagej fiji software, and normalised optical density was presented as a mean ± SD. In case of Tregs (FoxP3), results were presented as an absolute number of positive cells. Statistical analysis was performed using two‐tailed, unpaired Student's t ‐test with graphpad prism 8 software, n = 4.

Article Snippet: Human colorectal adenocarcinoma SW620 (ATCC ® CCL‐227; RRID:CVCL_0547) and HCT116 (ATCC ® CCL‐247; RRID:CVCL_0291), human breast cancer MCF7 (ATCC ® HTB‐22; RRID:CVCL_0031), human lung cancer A549 (ATCC ® CCL‐185; RRID:CVCL_JK07) and the mouse colon carcinoma cell line colon26 (ATCC ® CRL‐2639; RRID:CVCL_7255) were obtained from ATCC (Manassas, VA, USA).

Techniques: Immunohistochemical staining, Staining, Control, Software, Two Tailed Test

Journal: Molecular Oncology

Article Title: Immune modulation underpins the anti‐cancer activity of HDAC inhibitors

doi: 10.1002/1878-0261.12953

Figure Lengend Snippet: Treatment of colon26 tumours with CXD101 and anti‐PD‐1. (A) Schematic representation of the experiment with CXD101 in colon26 tumours (i). Balb/c mice were treated with orally administrated CXD101 (50 mg·kg −1 ; 5‐day on/2‐day off schedule) for 38 days or the vehicle‐only control. Additionally, group 3 was treated with anti‐mPD‐1 monotherapy administered intraperitoneally, twice weekly at a dose level of 5 mg·kg −1 . Group 4 received a combination of CXD101 with anti‐mPD‐1; n = 6; (ii) scatter plots of relative tumour volume of individual mice at day 15 (Student's t ‐test; * P < 0.05); n = 6; (iii) relative growth analysis of treated and nontreated tumours (Student's t ‐test of the values at day 15; * P < 0.05); n = 8 (iv) relative body weight representation of treated and nontreated mice; n = 6; (v) survival curves of treated and nontreated mice (log‐rank [Mantel–Cox] test; * P < 0.05); n = 6. (B) Schematic representation of mouse experiment with CXD101 in MC38 tumours (i). C57BL/6 mice were treated with orally administrated CXD101 (50 mg·kg −1 ; 5‐day on/2‐day off schedule) for 38 days ( n = 6) or the vehicle‐only control ( n = 8). Group 3 mice were treated with anti‐mPD‐1 monotherapy ( n = 8), administered intraperitoneally twice per week (10 mg·kg −1 ; days 1, 6, 11, 16, 21 and 26). Group 4 received a combination of CXD101 and anti‐mPD‐1; n = 6; (ii) scatter plots of relative tumour volume of individual mouse at day 13 (Student's t ‐test; * P < 0.05); (iii) relative tumour growth analysis of treated and nontreated mice presented as a mean value (Student's t ‐test of the values at day 13; * P < 0.05); (iv) relative body weight of treated and nontreated mice presented as a mean value; (v) survival curves of treated and nontreated mice (log‐rank (Mantel–Cox) test; * P < 0.05). (C) Model hypothesis for the effect of HDAC inhibition in regulating the immune response to tumours. It is proposed that inhibition of histone deacetylases induces expression of MHC genes and thereby increases antigen presentation, which together with the release of T cells from immune checkpoint inhibition with anti‐PD‐1 contributes to improved T‐cell engagement via MHC class I and improved tumour cell killing.

Article Snippet: Human colorectal adenocarcinoma SW620 (ATCC ® CCL‐227; RRID:CVCL_0547) and HCT116 (ATCC ® CCL‐247; RRID:CVCL_0291), human breast cancer MCF7 (ATCC ® HTB‐22; RRID:CVCL_0031), human lung cancer A549 (ATCC ® CCL‐185; RRID:CVCL_JK07) and the mouse colon carcinoma cell line colon26 (ATCC ® CRL‐2639; RRID:CVCL_7255) were obtained from ATCC (Manassas, VA, USA).

Techniques: Control, Inhibition, Expressing, Immunopeptidomics

Journal: Molecular Therapy Oncolytics

Article Title: Whole cell vaccination using immunogenic cell death by an oncolytic adenovirus is effective against a colorectal cancer model

doi: 10.1038/mto.2016.31

Figure Lengend Snippet: Schematic structure of adenoviral vectors and cytotoxic efficiencies of oncolytic adenoviruses in colorectal cancer (CRC) cells. ( a ) Schematic structure of adenoviral vectors. Schematic structures of Ad881 and Ad884 are shown. ITR, adenovirus inverted terminal repeat sequence; ψ, packaging signal; pA, polyadenylation signal; IRES, internal ribosome entry site; EGFP, enhanced green fluorescent protein; CMV, cytomegalovirus promoter; Mdk, midkine; HSV-TK, herpes simplex virus-thymidine kinase. ( b )/( c ) Cyotopathic assays using Ad884 in CRC cells and normal fibroblasts. CRC cell lines (DLD-1, CMT93, and CT26) and fibroblasts were infected with Ad884 ( b ) or Ad881 ( c ) at various multiplicity of infections (MOIs) (0, 0.1, 1, 10, 100, or 1,000). On day 8, cytotoxicity was assessed by the extent of crystal violet staining. ( d ) Virus progeny production in CRC cells. DLD-1, CMT93, and CT26 cells, as well as fibroblasts were infected with Ad881 or Ad884 at an MOI of 100. At 48 hours after infection, cells and media were harvested to determine the viral titer in transducing units by EGFP expression using flow cytometry. Virus production levels were normalized to the baseline value in fibroblasts. Data are representative of three independent experiments all yielding similar results. Ad881: black bar; Ad884: hatched bar. ( e ) Time-dependent cytotoxicity. DLD-1 (triangle) and CT26 (circle) cells (1 × 10 4 /well) were cultured as multiple replicates in 96-well plates and infected with Ad881 (closed) or Ad884 (open) at an MOI of 100. On the indicated days, the number of surviving cells was analyzed by a colorimetric method using Alamar blue. Data shown are the mean ± SD of triplicates.

Article Snippet: Mouse CRC cell lines Colon-26 (CT26) and CMT93 were purchased from RIKEN BRC Cell Bank (Ibaraki, Japan) and DS Pharma Biomedical (Osaka, Japan), respectively.

Techniques: Sequencing, Virus, Infection, Staining, Expressing, Flow Cytometry, Cell Culture

Journal: Molecular Therapy Oncolytics

Article Title: Whole cell vaccination using immunogenic cell death by an oncolytic adenovirus is effective against a colorectal cancer model

doi: 10.1038/mto.2016.31

Figure Lengend Snippet: Infection efficacy of the oncolytic adenovirus Ad881 in CT26 cells. Representative bright field (top) and enhanced green fluorescence protein (EGFP) fluorescence (bottom) images of CT26 cells that were left uninfected (left) or infected with Ad881 at an multiplicity of infection (MOI) of 10 (middle) or 1,000 (right).

Article Snippet: Mouse CRC cell lines Colon-26 (CT26) and CMT93 were purchased from RIKEN BRC Cell Bank (Ibaraki, Japan) and DS Pharma Biomedical (Osaka, Japan), respectively.

Techniques: Infection, Fluorescence

Journal: Molecular Therapy Oncolytics

Article Title: Whole cell vaccination using immunogenic cell death by an oncolytic adenovirus is effective against a colorectal cancer model

doi: 10.1038/mto.2016.31

Figure Lengend Snippet: Assessment of vaccine conditions. ( a ) The tumor formation rate in mice following subcutaneous injection with 1 × 10 5 Ad881-infected CT26 cells that had been infected for 10 hours at multiplicity of infections (MOIs) of 0, 10, 100, or 1,000. ( b ) The tumor formation rate in mice following subcutaneous injection of 1 × 10 4 or 1 × 10 5 Ad881-infected CT26 cells that had been infected for 24 hours at MOIs of 0 or 1,000.

Article Snippet: Mouse CRC cell lines Colon-26 (CT26) and CMT93 were purchased from RIKEN BRC Cell Bank (Ibaraki, Japan) and DS Pharma Biomedical (Osaka, Japan), respectively.

Techniques: Injection, Infection

Journal: Molecular Therapy Oncolytics

Article Title: Whole cell vaccination using immunogenic cell death by an oncolytic adenovirus is effective against a colorectal cancer model

doi: 10.1038/mto.2016.31

Figure Lengend Snippet: Assessment of vaccine efficiency. ( a ) Schema of the vaccination protocol used in this study. ( b ) The tumor rejection rate in mice that had been previously vaccinated with 1 × 10 4 of Ad881-infected (MOI: 0 or 1,000) CT26 cells or no previous treatment when these mice were injected with 1 × 10 4 uninfected CT26 as a challenge. ( c ) The tumor rejection rate following challenge with 1 × 10 5 uninfected CT26 cells in mice that were vaccinated with 1 × 10 4 Ad881-infected CT26 cells once or twice (repeatedly). MOI, multiplicity of infection.

Article Snippet: Mouse CRC cell lines Colon-26 (CT26) and CMT93 were purchased from RIKEN BRC Cell Bank (Ibaraki, Japan) and DS Pharma Biomedical (Osaka, Japan), respectively.

Techniques: Infection, Injection