ct26 cells  (ATCC)

Journal: Bioactive Materials

Article Title: Chronic inflammation-responsive hydrogel restores myeloid-T cell crosstalk to reinvigorate antitumor immunity against metastatic colorectal cancer

doi: 10.1016/j.bioactmat.2026.03.012

Figure Lengend Snippet: PGE2 blockade modulates immune cell phenotypes in antitumor resp onses. (A) Inflammatory gene expression across cancer types (GEPIA2 database). (B) Gene expression of Il1b , Cxcl8 , and Lif in colon adenocarcinoma (COAD) tumor tissue and normal tissue (GEPIA2 database). (C and D) Correlation between Ptgs2 and inflammatory genes in various cancers (C) and COAD (D) (TIMER 2.0). (E) Schematic of immune cells co-incubated with CXB treated tumor conditional medium (TCM) (Source material from BioRender). (F and G) Cell viability (F) and Cell cycle arrest (G) detection of CT26 tumor cells treated with gradient concentrations of CXB; n = 3. (H) PGE2 concentration in CT26 cell supernatants; n = 3. (I) The proportion of CD103 + DC within BMDCs after CXB treatments in vitro ; n = 3. (J and K) Maturation (J) and Antigen processing capability (K) on BMDCs; n = 3. (L – N) Flow charts of CD86 or CD206 expression on Raw 264.7 cells (L). Quantification of CD86 (M) and CD206 (N) expression on Raw 264.7 cells; n = 3. (O and P) Flow charts (O) and Quantification (P) of CD69 and CD137 expression on splenic T cells exposed to CXB-pretreated TCM; n = 3. (Q) IFN-γ secretion by T cells co-cultured with CXB-pretreated TCM; n = 3. Data are presented as mean ± SD, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Significance was calculated using One-way ANOVA.

Article Snippet: CT26 cells and Raw 264.7 were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Gene Expression, Incubation, Concentration Assay, In Vitro, Expressing, Cell Culture

Journal: Bioactive Materials

Article Title: Chronic inflammation-responsive hydrogel restores myeloid-T cell crosstalk to reinvigorate antitumor immunity against metastatic colorectal cancer

doi: 10.1016/j.bioactmat.2026.03.012

Figure Lengend Snippet: Sustained PGE2 blockade prompts immune activ ation. (A) Structure of hydrogel matrix and scheme of Gel-CXB preparation (Source material from BioRender). (B) Microstructure of the hydrogel. (C) Rheological evaluation of Gel-CXB. (D) CXB release from Gel-CXB in PBS or PBS containing 0.5 mM H 2 O 2 ; n = 3. (E and F) Flow chart (E) and Quantification (F) of CD103 + DC within BMDCs; n = 3. (G and H) Flow chart (G) and Heatmap (H) of costimulatory molecular expression on CD103 - DC, CD103 + DC, or total DC with different treatments; n = 3. (I and J) CXCL9 (I) and Costimulatory molecular expression (J) on cDC1; n = 3. (K – M) CD86 and CD206 expression (K), MHC-II expression (L), and Antigen processing capability (M) of BMDMs incubated with different TCM; n = 3. (N and O) CD69 (N) and CD137 (O) expression on CD8 + T cells co-incubated with different TCM; n = 3. (P) Scheme of Gel-CXB-regulated CT26 TME at different time points in vivo . (Q) Changes of several immune cells within TME at Day 1, 5, and 9; n = 3. (R) Tumor volume of mice treated with CXB alone or Gel-CXB in vivo ; n = 5. (S) CD137 expression on CD8 + T cells in vivo ; n = 3. Data are presented as mean ± SD, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Significance was calculated using One-way ANOVA.

Article Snippet: CT26 cells and Raw 264.7 were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Expressing, Incubation, In Vivo

Journal: Bioactive Materials

Article Title: Chronic inflammation-responsive hydrogel restores myeloid-T cell crosstalk to reinvigorate antitumor immunity against metastatic colorectal cancer

doi: 10.1016/j.bioactmat.2026.03.012

Figure Lengend Snippet: TRANS inhibits tumor growth and enhances local and systemic immune resp onses. (A) Scheme of GC, GCF, or TRANS preparation (Source material from BioRender). (B) Microstructure of GC and TRANS. (C) Experimental design for administration and immune cell analysis. (D) Tumor growth curves under different treatments; n = 5. (E) Tumor weight post-treatment; n = 5. (F – H) CD45 + leukocytes and CD11c + DCs (F), CD86 + M1 and CD206 + M2 macrophages (G), and Tumor-infiltrating CD8 + T cells (H) within TME; n = 5. (I – L) Mature DCs (I), CD8α + cDC1s (J), CD4 + and CD8 + T cells (K) and CD69 + CD8 + T cells (L) in lymph nodes; n = 5. (M – Q) CD11c + MHC II + DCs (M), CD8α + cDC1s (N), CD4 + and CD8 + T cells (O), CD69 + CD8 + T cells (P), and IFN-γ + CD8 + T cells (Q) in the spleen; n = 5. (R – T) CD8 + T cells (R), The ratio of CD8 + T /CD4 + T cells (S), and IFN-γ levels (T) in blood; n = 5. (U) IFN-γ + CD4 + T and IFN-γ + CD8 + T cells with ex vivo stimulation of PMA/ionomycin for 6 h; n = 3. (V) Apoptosis of CT26 cells co-incubated with splenic T cells for 24 h; n = 3. Data are presented as mean ± SD, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Significance was calculated using One-way ANOVA.

Article Snippet: CT26 cells and Raw 264.7 were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Cell Analysis, Ex Vivo, Incubation

Journal: Bioactive Materials

Article Title: Chronic inflammation-responsive hydrogel restores myeloid-T cell crosstalk to reinvigorate antitumor immunity against metastatic colorectal cancer

doi: 10.1016/j.bioactmat.2026.03.012

Figure Lengend Snippet: TRANS inhibits tumor metastasis and induces immune memory in vivo . (A) Experimental design for secondary tumor model. (B and C) Tumor volume curves of primary tumor (B) and secondary tumor (C) during different therapy; n = 5. (D and E) Statistical diagram (D) and flow charts (E) of T cells within secondary tumors; n = 5. (F) Immunofluorescence images of immune cell in primary tumor. (G) Schematic of lung metastasis tumor model and treatment regimen. Mice received subcutaneous and intravenous injections of CT26-Luc. (H – J) In vivo images (H), Primary tumor volume curves (I), and Average radiance in lungs (J) of CT26-Luc tumor-bearing mice; n = 5. (K – M) Lung image (K), Lung metastasis foci counts and weights (L), and H&E staining of lungs (M) from CT26-Luc tumor-bearing mice; n = 5. (N) Schematic of liver metastasis tumor model and treatment regimen. Mice received subcutaneous CT26 tumor and splenic CT26-Luc injections. (O and P) In vivo imaging (O) and Individual radiance in livers (P) of CT26-Luc tumor-bearing mice; n = 10. (Q – S) Live images (Q), Liver weights (R), and H&E staining images of livers (S) from PBS- or TRANS-treated mice; n = 5. (T) Scheme of tumor rechallenge model. (U) Tumor changes in mice rechallenged with CT26 or 4T1; n = 9. (V) Central memory (T CM ) and effector memory (T EM ) gated on CD8 + T cells; n = 5. Data are presented as mean ± SD, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Significance was calculated using One-way ANOVA.

Article Snippet: CT26 cells and Raw 264.7 were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: In Vivo, Immunofluorescence, Staining, In Vivo Imaging

Journal: bioRxiv

Article Title: Lung-targeted cytokine-coding RNA-lipoplexes induce T and NK cell-mediated anti-tumor immune response

doi: 10.64898/2026.05.06.723126

Figure Lengend Snippet: ( A-D ) Luc–encoding RNA formulated as RNA-LPX was administered i.v. to non-tumor bearing (A, B; n=3) or CT26 metastases-bearing (C, D) BALB/c mice (n=3-9). ( A ) BLI of total body, ( B ) normalized organ signal from explanted organs. Data were analyzed by ordinary one-way ANOVA and Tukey’s test for multiple comparisons. ( C ) Kinetics of BLI of the lung signal in metastases-bearing mice, significance was determined by mixed-effects analysis with Geisser-Greenhouse correction and Tukey’s test for multiple comparisons. ( D ) Luc RNA, luc protein and CD31 (PECAM-1) were detected via RNAscope and immunohistochemistry on consecutive sections 1, 6 or 24 hours post injection, scale bar: 100 µm, tumor tissue is indicated with dashed line. ( E ) Cytokine quantification in lung after the indicated time points. ( F ) Cytokine fold increase in lung normalized to spleen at 6 hours after injection. ( G ) FDG positron emission tomography (PET) imaging 24 h after cytokine RNA mix injection into naïve mice, ( H ) ex vivo measurement of FDG uptake. Significance was determined using a two-tailed t-test for unpaired samples.

Article Snippet: Colon Carcinoma cell line CT26 was purchased from ATCC (CT26: CRL-2638, lot no. 58494154, female).

Techniques: RNAscope, Immunohistochemistry, Injection, Positron Emission Tomography, Imaging, Ex Vivo, Two Tailed Test

Journal: bioRxiv

Article Title: Lung-targeted cytokine-coding RNA-lipoplexes induce T and NK cell-mediated anti-tumor immune response

doi: 10.64898/2026.05.06.723126

Figure Lengend Snippet: ( A ) Experimental design: BALB/c mice (n=15 per group) were injected i.v. with CT26 tumor cells; subsequently, cytokine RNA mix or irrelevant RNA was administered i.v. as RNA-LPX twice per week for a total of 8 injections from d3 to d27. Survivor mice from the cytokine RNA mix-treated group (n=5) compared to naïve BALB/c mice (n=10) were re-challenged with CT26 tumor cells. ( B ) Survival after the initial i.v. CT26 tumor cell inoculation. ( C ) Survival after re-challenge. Significance was determined via Mantel-Cox logrank. ( D-F ) BALB/c mice (n=5 per group) were injected i.v. with CT26 tumor cells, RNA-LPX was administered i.v. at d3, d6 and d10 p.t.i., mice were sacrificed at d11, lungs were collected and analyzed via flow cytometry. Significance for pairwise comparisons was determined by unpaired two-tailed t-test. Flow cytometric analysis of ( D ) tumor burden, ( E ) CD8 + T and NK cells and ( F ) CD4 + Foxp3 + CD25 + T reg . ( G ) Functional analysis: after tumor cell inoculation, mice were treated with RNA-LPX at d3, d6, d10 and d13 p.t.i., mice were sacrificed at d14 for an ex vivo stimulation assay with PMA/Ionomycin ( H ) qRT-PCR was done to determine fold-change expression of cytokine RNA mix-treated normalized to irrelevant RNA LPX-treated lung samples (n=5 per group indicated in rows) for the indicated cytokines and chemokines.

Article Snippet: Colon Carcinoma cell line CT26 was purchased from ATCC (CT26: CRL-2638, lot no. 58494154, female).

Techniques: Injection, Flow Cytometry, Two Tailed Test, Functional Assay, Ex Vivo, Quantitative RT-PCR, Expressing

Journal: bioRxiv

Article Title: Lung-targeted cytokine-coding RNA-lipoplexes induce T and NK cell-mediated anti-tumor immune response

doi: 10.64898/2026.05.06.723126

Figure Lengend Snippet: Experimental design: BALB/c mice (n=5 per group) were injected i.v. with CT26 tumor cells; cytokine RNA mix or irrelevant RNA was administered i.v. at d3, d6 and d10 post tumor cell injection, mice were sacrificed at d11, lungs were collected and pooled at same ratios before sorting of CD45 + cells. CD45 + cells were subjected to scRNAseq. ( A,B ) Uniform manifold approximation and projection (UMAP) of 22 assigned clusters ( A ) and cell type frequencies ( B ) of CD45 + cells isolated from the lungs of cytokine RNA mix-treated versus irrelevant RNA-treated mice. ( C ) Effector function visualized as bubble plot. ( D ) Selected differentially expressed genes in cytokine RNA mix-treated versus irrelevant RNA-treated samples are shown as average log2 fold change. Note only significantly expressed values (p ≤ 0.05) are shown in colouring (blue = downregulated, red = upregulated), non-significant values are set to “0”/white.

Article Snippet: Colon Carcinoma cell line CT26 was purchased from ATCC (CT26: CRL-2638, lot no. 58494154, female).

Techniques: Injection, Isolation

Journal: bioRxiv

Article Title: Lung-targeted cytokine-coding RNA-lipoplexes induce T and NK cell-mediated anti-tumor immune response

doi: 10.64898/2026.05.06.723126

Figure Lengend Snippet: ( A ) Experimental design for ( B-C; CT26 tumor model) and ( E; CT26 B2M k.o. tumor model); n=15 BALB/c mice per group. Depletion or blocking antibody treatment was started 2 days prior to RNA-LPX treatment to ensure depletion before treatment start. ( B, C ) Survival according to termination criteria. ( D ) Experimental design and survival in CT26B2M k.o. or CT26gp70 k.o. tumor cells i.v. tumor model, n=15 mice per group. ( E ) BALB/c mice injected i.v. with CT26B2M k.o and depletion/neutralization antibodies were applied as shown in ( A ). Note that data in ( E ) were generated within the same experiment, irrelevant RNA + isotype mix as well as cytokine mix RNA + isotype mix refers to the same groups in all 3 plots. Survival was analyzed via Mantel-Cox logrank test. For ( B ) and ( C ), groups 1 and 2 were furthermore compared via logrank test with emphasis on early and late differences, ( B ) ## Logrank test with emphasis on late differences ((rho=0, lambda=1): group 1 vs 2: p=0,00235; ( C ) # Logrank test with emphasis on early differences (rho=1, lambda=0): group 1 vs 2: p=0,0378.

Article Snippet: Colon Carcinoma cell line CT26 was purchased from ATCC (CT26: CRL-2638, lot no. 58494154, female).

Techniques: Blocking Assay, Injection, Neutralization, Generated