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ATCC capan1 cells
Capan1 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 97/100, based on 1687 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/capan1+cells/Capan-1/pm41984591-586-0-2
Average 97 stars, based on 1687 article reviews

capan1 cells - by Bioz Stars, 2026-10

97/100 stars

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Cell Culture:

Article Title: Regulatory elements in the Sox9 locus license the initiation of pancreatic ductal adenocarcinoma.
Article Snippet: PANC-1 cells (ATCC; CRL-1469) were cultured in DMEM/GlutaMax media (Gibco, 31966-047) supplemented with 10% fetal bovine serum (FBS, Gibco, 17964671) and 1% penicillin-streptomycin (PenStrep, Gibco, 15-140-122). .. Capan1 cells (ATCC, HTB-79) were cultured in RPMI 1640/Glutamax media (Gibco, 61870010) supplemented with 10% FBS and 1% PenStrep. ..

Article Title: Guggulsterone decreases proliferation and metastatic behavior of pancreatic cancer cells by modulating JAK/STAT and Src/FAK signaling
Article Snippet: Horseradish peroxidase conjugated anti-mouse and anti-rabbit IgG were procured from GE Healthcare Biosciences (Uppsala, Sweden) and FITC-conjugated anti-mouse IgG was obtained from Invitrogen (California, U.S.A.). .. 2.2 Cell lines and cell culture conditions The human PC highly aggressive cell lines- CD18/HPAF and Capan1 cells were procured from American Type Culture Collection (ATCC), and cultured in Dulbecco’s Modified Eagles Medium (DMEM, Sigma Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum and antibiotics (100 μg/mL penicillin and streptomycin) [ 27 ]. ..

Article Title: Guggulsterone decreases proliferation and metastatic behavior of pancreatic cancer cells by modulating JAK/STAT and Src/FAK signaling
Article Snippet: Horseradish peroxidase conjugated anti-mouse and anti-rabbit IgG were procured from GE Healthcare Biosciences (Uppsala, Sweden) and FITC-conjugated anti-mouse IgG was obtained from Invitrogen (California, U.S.A.). .. The human PC highly aggressive cell lines- CD18/HPAF and Capan1 cells were procured from American Type Culture Collection (ATCC), and cultured in Dulbecco’s Modified Eagles Medium (DMEM, Sigma Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum and antibiotics (100 μg/mL penicillin and streptomycin) [ 27 ]. ..

Stable Transfection:

Article Title: ELR+ CXC chemokines and oncogenic Ras-mediated tumorigenesis
Article Snippet: SW1990 (American Type Culture Collection) cells were stably transduced with pCMVNeo-TET and pSUPER-RETRO-PURO-TETO-KRas G12D shRNA similar to approaches described previously ( 24 ) to create the cell line SW1990 DOX KRas G12D shRNA. .. HRas G12V HEK-HT and AsPC1, Panc1, SW1990 and Capan1 cells (American Type Culture Collection) were stably infected with retroviruses derived from pSUPER-RETRO-PURO scramble or hCXCL1 shRNA-1 using a protocol described previously ( 30 ). ..

Infection:

Derivative Assay:

shRNA:

Modification:

Article Title: Guggulsterone decreases proliferation and metastatic behavior of pancreatic cancer cells by modulating JAK/STAT and Src/FAK signaling
Article Snippet: Horseradish peroxidase conjugated anti-mouse and anti-rabbit IgG were procured from GE Healthcare Biosciences (Uppsala, Sweden) and FITC-conjugated anti-mouse IgG was obtained from Invitrogen (California, U.S.A.). .. The human PC highly aggressive cell lines- CD18/HPAF and Capan1 cells were procured from American Type Culture Collection (ATCC), and cultured in Dulbecco’s Modified Eagles Medium (DMEM, Sigma Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum and antibiotics (100 μg/mL penicillin and streptomycin) [ 27 ]. ..

Image Search Results

Limited T cell persistence restricts durable tumor control in a dual target setting (1A) Illustrations of the CARs targeting PSCA (top) and MUC1 (bottom) used to generate CAR T cells. (1B) PSCA (left) and MUC1 (right) CAR expression in C.P and C.M T cells measured by flow cytometry 3 days post-transduction. Non-transduced (NT) T cells used as negative controls (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (1C) Detection of PSCA (top) and MUC1 (bottom) expression by CAPAN1 tumor cells measured using flow cytometry. (1D) Cytolytic activity of C.P T cells against 293T (PSCA negative control) and CAPAN1 tumor cells measured using Cr-release assay at the effector to target ratio (E:T) of 20:1. NT cells used as controls (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (1E) Anti-tumor activity of C.M T cells against 293T and CAPAN1 cells measured using Cr-release assay at the effector to target ratio (E:T) of 20:1 (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (1F) CAPAN1 tumor luminescence measured by IVIS imaging in a long-term co-culture assay with CAR T cells in G-Rex 6-well plates. Untreated (tumor-only) condition used as control (one-way Anova on day 27, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (1G) PSCA (top) and MUC1 (bottom) expression by CAPAN1 cells from the co-culture experiment in (1F), measured on day 20 by flow cytometry. All T cell treated conditions normalized to the control (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (1H) CAPAN1 luminescence in C.M, C.P, and C.P+C.M treated conditions in (1F) after re-treatment with respective CAR T cells on day 27 (t-tests on day 35, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001).

Journal: bioRxiv

Article Title: IL-7 armed binary CAR T cell strategy to augment potency against solid tumors

doi: 10.1101/2025.06.25.661524

Figure Lengend Snippet: Limited T cell persistence restricts durable tumor control in a dual target setting (1A) Illustrations of the CARs targeting PSCA (top) and MUC1 (bottom) used to generate CAR T cells. (1B) PSCA (left) and MUC1 (right) CAR expression in C.P and C.M T cells measured by flow cytometry 3 days post-transduction. Non-transduced (NT) T cells used as negative controls (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (1C) Detection of PSCA (top) and MUC1 (bottom) expression by CAPAN1 tumor cells measured using flow cytometry. (1D) Cytolytic activity of C.P T cells against 293T (PSCA negative control) and CAPAN1 tumor cells measured using Cr-release assay at the effector to target ratio (E:T) of 20:1. NT cells used as controls (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (1E) Anti-tumor activity of C.M T cells against 293T and CAPAN1 cells measured using Cr-release assay at the effector to target ratio (E:T) of 20:1 (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (1F) CAPAN1 tumor luminescence measured by IVIS imaging in a long-term co-culture assay with CAR T cells in G-Rex 6-well plates. Untreated (tumor-only) condition used as control (one-way Anova on day 27, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (1G) PSCA (top) and MUC1 (bottom) expression by CAPAN1 cells from the co-culture experiment in (1F), measured on day 20 by flow cytometry. All T cell treated conditions normalized to the control (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (1H) CAPAN1 luminescence in C.M, C.P, and C.P+C.M treated conditions in (1F) after re-treatment with respective CAR T cells on day 27 (t-tests on day 35, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001).

Article Snippet: For co-culture experiments, GFP/FFLuc expressing CAPAN1 tumor cells (1×10 6 cells) were inoculated into 3D algimatrix bioscaffold (Thermo Fisher Scientific, Inc., Waltham, MA) and cultured in 6-well G-Rex devices (Wilson Wolf Manufacturing, New Brighton, MN).

Techniques: Control, Expressing, Flow Cytometry, Transduction, Activity Assay, Negative Control, Release Assay, Imaging, Co-culture Assay, Co-Culture Assay

Engineering CAR T cells to secrete and efficiently utilize IL-7 cytokine (2A) Illustrations of the IL-7 cytokine construct containing mOrange fluorescent protein for transgene detection by flow cytometry (top) and detection of both the PSCA CAR and IL-7 transgene expression in C.P7 T cells from a representative donor after serial transduction (bottom). (2B) Summary data indicating PSCA CAR and IL-7 double-transduced T cells compared to NT cells (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (2C) Production of IL-7 by C.P7 cells after stimulation with irradiated K562 cells engineered to express PSCA, measuring using ELISA (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (2D) Diagram illustrating the IL-7Rα construct containing GFP for transgene detection (top) and flow cytometry data for a representative donor demonstrating the expression of both the MUC1 CAR and the IL-7Rα transgenes in C.M7R T cells. (2E) Summary data comparing IL-7Rα detection by flow cytometry in NT, C.M, and C.M7R cells (one-way ANOVA, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (2F) Quantification of C.M and C.M7R T cells using trypan blue exclusion during culture with irradiated CAPAN1 tumor cells in presence or absence of recombinant IL-7 cytokine (t-tests on day 8, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001).

Journal: bioRxiv

Article Title: IL-7 armed binary CAR T cell strategy to augment potency against solid tumors

doi: 10.1101/2025.06.25.661524

Figure Lengend Snippet: Engineering CAR T cells to secrete and efficiently utilize IL-7 cytokine (2A) Illustrations of the IL-7 cytokine construct containing mOrange fluorescent protein for transgene detection by flow cytometry (top) and detection of both the PSCA CAR and IL-7 transgene expression in C.P7 T cells from a representative donor after serial transduction (bottom). (2B) Summary data indicating PSCA CAR and IL-7 double-transduced T cells compared to NT cells (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (2C) Production of IL-7 by C.P7 cells after stimulation with irradiated K562 cells engineered to express PSCA, measuring using ELISA (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (2D) Diagram illustrating the IL-7Rα construct containing GFP for transgene detection (top) and flow cytometry data for a representative donor demonstrating the expression of both the MUC1 CAR and the IL-7Rα transgenes in C.M7R T cells. (2E) Summary data comparing IL-7Rα detection by flow cytometry in NT, C.M, and C.M7R cells (one-way ANOVA, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (2F) Quantification of C.M and C.M7R T cells using trypan blue exclusion during culture with irradiated CAPAN1 tumor cells in presence or absence of recombinant IL-7 cytokine (t-tests on day 8, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001).

Techniques: Construct, Flow Cytometry, Expressing, Transduction, Irradiation, Enzyme-linked Immunosorbent Assay, Recombinant

IL-7 engineered C.P7 T cells support the expansion of C.M7R overexpressing IL-7R α (3A) Growth of C.M7R T cells in cell culture medium, and conditioned medium collected from C.P or C.P7 T cells stimulated with irradiated CAPAN1 tumor cells. Cells were quantified by manual counting (t-tests on day 5, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (3B) Total cell numbers in irradiated CAPAN1-stimulation co-cultures of the indicated T cell types (t-tests on day 14, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (3C) Differentiation of C.P7 and C.M7R T cells based on GFP (surrogate for transgenic IL-7Rα) using flow cytometry to quantify the proportion of these cells in the C.P7+C.M7R condition at the start and the end of the stimulation experiment in (3B) (t-tests on day 14, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (3D) Flow cytometric quantification of C.M7R T cells cultured with NT (control), C.P, or C.P7 T cells pre-stimulated with irradiated CAPAN1. Arrows indicate time points at which additional pre-stimulated C.P7 T cells were added. Dashed lines indicate conditions which were monitored for an additional time point without the additional dose of pre-stimulated C.P7 T cells (t-tests at indicated time-points, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001).

Journal: bioRxiv

Article Title: IL-7 armed binary CAR T cell strategy to augment potency against solid tumors

doi: 10.1101/2025.06.25.661524

Figure Lengend Snippet: IL-7 engineered C.P7 T cells support the expansion of C.M7R overexpressing IL-7R α (3A) Growth of C.M7R T cells in cell culture medium, and conditioned medium collected from C.P or C.P7 T cells stimulated with irradiated CAPAN1 tumor cells. Cells were quantified by manual counting (t-tests on day 5, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (3B) Total cell numbers in irradiated CAPAN1-stimulation co-cultures of the indicated T cell types (t-tests on day 14, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (3C) Differentiation of C.P7 and C.M7R T cells based on GFP (surrogate for transgenic IL-7Rα) using flow cytometry to quantify the proportion of these cells in the C.P7+C.M7R condition at the start and the end of the stimulation experiment in (3B) (t-tests on day 14, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (3D) Flow cytometric quantification of C.M7R T cells cultured with NT (control), C.P, or C.P7 T cells pre-stimulated with irradiated CAPAN1. Arrows indicate time points at which additional pre-stimulated C.P7 T cells were added. Dashed lines indicate conditions which were monitored for an additional time point without the additional dose of pre-stimulated C.P7 T cells (t-tests at indicated time-points, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001).

Techniques: Cell Culture, Irradiation, Transgenic Assay, Flow Cytometry, Control

Dual-targeted binary T cells exhibit enhanced anti-tumor activity and persistence (4A) Representative bioluminescent images of GFP/FFLuc expressing CAPAN1-seeded Algimatrix cultures in the long-term co-culture assay using 6-well G-Rex devices. Two replicates shown for each condition (4B) Quantification of bioluminescence signal from the CAPAN-1 cells [illustrated in (4A)] during the co-culture experiment under indicated treatment conditions (t-tests on day 20, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (4C) Live T cell counts using flow cytometry on day 20 for the different treatment conditions in (4B) (one-way ANOVA, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001).

Journal: bioRxiv

Article Title: IL-7 armed binary CAR T cell strategy to augment potency against solid tumors

doi: 10.1101/2025.06.25.661524

Figure Lengend Snippet: Dual-targeted binary T cells exhibit enhanced anti-tumor activity and persistence (4A) Representative bioluminescent images of GFP/FFLuc expressing CAPAN1-seeded Algimatrix cultures in the long-term co-culture assay using 6-well G-Rex devices. Two replicates shown for each condition (4B) Quantification of bioluminescence signal from the CAPAN-1 cells [illustrated in (4A)] during the co-culture experiment under indicated treatment conditions (t-tests on day 20, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (4C) Live T cell counts using flow cytometry on day 20 for the different treatment conditions in (4B) (one-way ANOVA, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001).

Techniques: Activity Assay, Expressing, Co-culture Assay, Co-Culture Assay, Flow Cytometry

Binary T cells demonstrate superior anti-tumor activity against pancreatic spheroids (5A) Representative bioluminescent images of GFP/FFLuc expressing CAPAN1 tumor cells generated spheroids during long-term co-culture. Three replicates illustrated for each condition. (5B) Quantitate bioluminescence data for the CAPAN1 spheroids in absence of treatment or during co-culture with indicated T cell types (t-tests on day 21, n=9, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (5C) Live T cell counts for the different treatment conditions in (5B) obtained using flow cytometry on day 14 (t-tests, n=4, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001).

Journal: bioRxiv

Article Title: IL-7 armed binary CAR T cell strategy to augment potency against solid tumors

doi: 10.1101/2025.06.25.661524

Figure Lengend Snippet: Binary T cells demonstrate superior anti-tumor activity against pancreatic spheroids (5A) Representative bioluminescent images of GFP/FFLuc expressing CAPAN1 tumor cells generated spheroids during long-term co-culture. Three replicates illustrated for each condition. (5B) Quantitate bioluminescence data for the CAPAN1 spheroids in absence of treatment or during co-culture with indicated T cell types (t-tests on day 21, n=9, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (5C) Live T cell counts for the different treatment conditions in (5B) obtained using flow cytometry on day 14 (t-tests, n=4, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001).

Techniques: Activity Assay, Expressing, Generated, Co-Culture Assay, Flow Cytometry

Limited T cell persistence restricts durable tumor control in a dual target setting. (A) Illustrations of the CARs targeting PSCA (top) and MUC1 (bottom) used to generate CAR T cells. (B) PSCA (left) and MUC1 (right) CAR expression in C.P and C.M T cells measured by flow cytometry 3 days post-transduction. Non-transduced (NT) T cells used as negative controls (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (C) Detection of PSCA (top) and MUC1 (bottom) expression by CAPAN1 tumor cells measured using flow cytometry. (D) Cytolytic activity of C.P T cells against 293T (PSCA negative control) and CAPAN1 tumor cells measured using 51 Cr-release assay at the effector to target ratio (E:T) of 20:1. NT cells used as controls (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (E) Anti-tumor activity of C.M T cells against 293T and CAPAN1 cells measured using 51 Cr-release assay at the effector to target ratio (E:T) of 20:1 (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (F) CAPAN1 tumor luminescence measured by IVIS imaging in a long-term co-culture assay with CAR T cells in G-Rex 6-well plates. Untreated (tumor-only) condition used as control (one-way Anova on day 27, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (G) PSCA (top) and MUC1 (bottom) expression by CAPAN1 cells from the co-culture experiment in (1F), measured on day 20 by flow cytometry. All T cell treated conditions normalized to the control (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (H) CAPAN1 luminescence in C.M, C.P, and C.P+C.M treated conditions in (1F) after re-treatment with respective CAR T cells on day 27 (t-tests on day 35, n=3, ns, no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001).

Journal: Frontiers in Immunology

Article Title: IL-7 armed binary CAR T cell strategy to augment potency against solid tumors

doi: 10.3389/fimmu.2025.1618404

Figure Lengend Snippet: Limited T cell persistence restricts durable tumor control in a dual target setting. (A) Illustrations of the CARs targeting PSCA (top) and MUC1 (bottom) used to generate CAR T cells. (B) PSCA (left) and MUC1 (right) CAR expression in C.P and C.M T cells measured by flow cytometry 3 days post-transduction. Non-transduced (NT) T cells used as negative controls (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (C) Detection of PSCA (top) and MUC1 (bottom) expression by CAPAN1 tumor cells measured using flow cytometry. (D) Cytolytic activity of C.P T cells against 293T (PSCA negative control) and CAPAN1 tumor cells measured using 51 Cr-release assay at the effector to target ratio (E:T) of 20:1. NT cells used as controls (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (E) Anti-tumor activity of C.M T cells against 293T and CAPAN1 cells measured using 51 Cr-release assay at the effector to target ratio (E:T) of 20:1 (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (F) CAPAN1 tumor luminescence measured by IVIS imaging in a long-term co-culture assay with CAR T cells in G-Rex 6-well plates. Untreated (tumor-only) condition used as control (one-way Anova on day 27, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (G) PSCA (top) and MUC1 (bottom) expression by CAPAN1 cells from the co-culture experiment in (1F), measured on day 20 by flow cytometry. All T cell treated conditions normalized to the control (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (H) CAPAN1 luminescence in C.M, C.P, and C.P+C.M treated conditions in (1F) after re-treatment with respective CAR T cells on day 27 (t-tests on day 35, n=3, ns, no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001).

Article Snippet: The cell lines CAPAN1, K562, and 293T were obtained from the American Type Culture Collection (Rockville, MD) and were grown in Dulbecco’s modified eagle medium (DMEM, GE Healthcare Life Sciences, Pittsburgh, PA) supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Hyclone, Waltham, MA) and 2 mM L-GlutaMAX (Gibco BRL Life Technologies, Inc., Gaithersburg, MD).

Techniques: Control, Expressing, Flow Cytometry, Transduction, Activity Assay, Negative Control, Release Assay, Imaging, Co-culture Assay, Co-Culture Assay

Engineering CAR T cells to secrete and efficiently utilize IL-7 cytokine (A). Illustrations of the IL-7 cytokine construct containing mOrange fluorescent protein for transgene detection by flow cytometry (top) and detection of both the PSCA CAR and IL-7 transgene expression in C.P7 T cells from a representative donor after serial transduction (bottom). (B) Summary data indicating PSCA CAR and IL-7 double-transduced T cells compared to NT cells (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (C) Production of IL-7 by C.P7 cells after stimulation with irradiated K562 cells engineered to express PSCA, measuring using ELISA (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (D) Diagram illustrating the IL-7Rα construct containing GFP for transgene detection (top) and flow cytometry data for a representative donor demonstrating the expression of both the MUC1 CAR and the IL-7Rα transgenes in C.M7R T cells. (E) Summary data comparing IL-7Rα detection by flow cytometry in NT, C.M, and C.M7R cells (one-way ANOVA, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (F) Quantification of C.M and C.M7R T cells using trypan blue exclusion during culture with irradiated CAPAN1 tumor cells in presence or absence of recombinant IL-7 cytokine (t-tests on day 8, n=3, ns, no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001).

Journal: Frontiers in Immunology

Article Title: IL-7 armed binary CAR T cell strategy to augment potency against solid tumors

doi: 10.3389/fimmu.2025.1618404

Figure Lengend Snippet: Engineering CAR T cells to secrete and efficiently utilize IL-7 cytokine (A). Illustrations of the IL-7 cytokine construct containing mOrange fluorescent protein for transgene detection by flow cytometry (top) and detection of both the PSCA CAR and IL-7 transgene expression in C.P7 T cells from a representative donor after serial transduction (bottom). (B) Summary data indicating PSCA CAR and IL-7 double-transduced T cells compared to NT cells (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (C) Production of IL-7 by C.P7 cells after stimulation with irradiated K562 cells engineered to express PSCA, measuring using ELISA (t-tests, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (D) Diagram illustrating the IL-7Rα construct containing GFP for transgene detection (top) and flow cytometry data for a representative donor demonstrating the expression of both the MUC1 CAR and the IL-7Rα transgenes in C.M7R T cells. (E) Summary data comparing IL-7Rα detection by flow cytometry in NT, C.M, and C.M7R cells (one-way ANOVA, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (F) Quantification of C.M and C.M7R T cells using trypan blue exclusion during culture with irradiated CAPAN1 tumor cells in presence or absence of recombinant IL-7 cytokine (t-tests on day 8, n=3, ns, no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001).

Techniques: Construct, Flow Cytometry, Expressing, Transduction, Irradiation, Enzyme-linked Immunosorbent Assay, Recombinant

IL-7 engineered C.P7 T cells support the expansion of C.M7R overexpressing IL-7Rα. (A) Growth of C.M7R T cells in cell culture medium, and conditioned medium collected from C.P or C.P7 T cells stimulated with irradiated CAPAN1 tumor cells. Cells were quantified by manual counting (t-tests on day 5, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (B) Total cell numbers in irradiated CAPAN1-stimulation co-cultures of the indicated T cell types (t-tests on day 14, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (C) Differentiation of C.P7 and C.M7R T cells based on GFP (surrogate for transgenic IL-7Rα) using flow cytometry to quantify the proportion of these cells in the C.P7+C.M7R condition at the start and the end of the stimulation experiment in (3B) (t-tests on day 14, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (D) Flow cytometric quantification of C.M7R T cells cultured with NT (control), C.P, or C.P7 T cells pre-stimulated with irradiated CAPAN1. Arrows indicate time points at which additional pre-stimulated C.P7 T cells were added. Dashed lines indicate conditions which were monitored for an additional time point without the additional dose of pre-stimulated C.P7 T cells (t-tests at indicated time-points, n=3, ns, no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001).

Journal: Frontiers in Immunology

Article Title: IL-7 armed binary CAR T cell strategy to augment potency against solid tumors

doi: 10.3389/fimmu.2025.1618404

Figure Lengend Snippet: IL-7 engineered C.P7 T cells support the expansion of C.M7R overexpressing IL-7Rα. (A) Growth of C.M7R T cells in cell culture medium, and conditioned medium collected from C.P or C.P7 T cells stimulated with irradiated CAPAN1 tumor cells. Cells were quantified by manual counting (t-tests on day 5, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (B) Total cell numbers in irradiated CAPAN1-stimulation co-cultures of the indicated T cell types (t-tests on day 14, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (C) Differentiation of C.P7 and C.M7R T cells based on GFP (surrogate for transgenic IL-7Rα) using flow cytometry to quantify the proportion of these cells in the C.P7+C.M7R condition at the start and the end of the stimulation experiment in (3B) (t-tests on day 14, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (D) Flow cytometric quantification of C.M7R T cells cultured with NT (control), C.P, or C.P7 T cells pre-stimulated with irradiated CAPAN1. Arrows indicate time points at which additional pre-stimulated C.P7 T cells were added. Dashed lines indicate conditions which were monitored for an additional time point without the additional dose of pre-stimulated C.P7 T cells (t-tests at indicated time-points, n=3, ns, no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001).

Techniques: Cell Culture, Irradiation, Transgenic Assay, Flow Cytometry, Control

Dual-targeted binary T cells exhibit enhanced anti-tumor activity and persistence. (A) Representative bioluminescent images of GFP/FFLuc expressing CAPAN1-seeded Algimatrix cultures in the long-term co-culture assay using 6-well G-Rex devices. Two replicates shown for each condition (B) Quantification of bioluminescence signal from the CAPAN-1 cells [illustrated in (4A)] during the co-culture experiment under indicated treatment conditions (t-tests on day 20, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (C) Live T cell counts using flow cytometry on day 20 for the different treatment conditions in (B) (one-way ANOVA, n=3, ns, no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001).

Journal: Frontiers in Immunology

Article Title: IL-7 armed binary CAR T cell strategy to augment potency against solid tumors

doi: 10.3389/fimmu.2025.1618404

Figure Lengend Snippet: Dual-targeted binary T cells exhibit enhanced anti-tumor activity and persistence. (A) Representative bioluminescent images of GFP/FFLuc expressing CAPAN1-seeded Algimatrix cultures in the long-term co-culture assay using 6-well G-Rex devices. Two replicates shown for each condition (B) Quantification of bioluminescence signal from the CAPAN-1 cells [illustrated in (4A)] during the co-culture experiment under indicated treatment conditions (t-tests on day 20, n=3, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (C) Live T cell counts using flow cytometry on day 20 for the different treatment conditions in (B) (one-way ANOVA, n=3, ns, no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001).

Techniques: Activity Assay, Expressing, Co-culture Assay, Co-Culture Assay, Flow Cytometry

Binary T cells demonstrate superior anti-tumor activity against pancreatic spheroids. (A) Representative bioluminescent images of GFP/FFLuc expressing CAPAN1 tumor cells generated spheroids during long-term co-culture. Three replicates illustrated for each condition. (B) Quantitate bioluminescence data for the CAPAN1 spheroids in absence of treatment or during co-culture with indicated T cell types (t-tests on day 21, n=9, ns, no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (C) Live T cell counts for the different treatment conditions in (B) obtained using flow cytometry on day 14 (t-tests, n=4, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001).

Journal: Frontiers in Immunology

Article Title: IL-7 armed binary CAR T cell strategy to augment potency against solid tumors

doi: 10.3389/fimmu.2025.1618404

Figure Lengend Snippet: Binary T cells demonstrate superior anti-tumor activity against pancreatic spheroids. (A) Representative bioluminescent images of GFP/FFLuc expressing CAPAN1 tumor cells generated spheroids during long-term co-culture. Three replicates illustrated for each condition. (B) Quantitate bioluminescence data for the CAPAN1 spheroids in absence of treatment or during co-culture with indicated T cell types (t-tests on day 21, n=9, ns, no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001). (C) Live T cell counts for the different treatment conditions in (B) obtained using flow cytometry on day 14 (t-tests, n=4, ns = no significant difference, *p<.05, **p<.01, ***p<.001, ****p<.0001).

Techniques: Activity Assay, Expressing, Generated, Co-Culture Assay, Flow Cytometry

Fig. 3 | JUNB-HDAC1 complex represses inﬂammatory signals and cJUN. a Heatmap showing expression of cytokines present in the core enrichment of the gene sets shown in Supplementary Fig. 3a–d, for JUNB silencing (siJUNB; n = 3 biological replicates) versus control siRNA (siCtrl; n = 2 biological replicates) in CAPAN1 cells. Cell color indicates z score. b qRT-PCR analysis for indicated target genes in siJUNB conditions (red), normalized to siCtrl (gray), in CAPAN1. Relative mRNA expression with mean ± s.d. shown. n = 3 biological replicates. Two-tailed Student’s t-test with Welch’s correction. Coverage of JUNB ChIP-seq data in CAPAN121, as well as publicly available H3K27ac25 data, for loci of cJUN (c), IL1A/B (d), and CXCL9/10/11 (e). ChIP-qPCR validation regions are indicated. f Gene set enrichment analysis for curated signatures (C2) of the Molecular Signature Data- base (MSigDB) for siJUNB versus siCtrl in CAPAN1 cells. Normalized enrichment score (NES) and FDR q value are indicated. Immunoblot for JUNB, HDAC1, and β- actin after JUNB pulldown, IgG isotype control or input in CAPAN1 (g) and CAPAN2 (h). n = 3 biological replicates. i, j, ChIP-qPCR for regions indicated in (c–e), showing signal relative to input for JUNB (i) and HDAC1 (j) pulldown with

Journal: Nature Communications

Article Title: Spatial tumor immune heterogeneity facilitates subtype co-existence and therapy response in pancreatic cancer

doi: 10.1038/s41467-024-55330-7

Figure Lengend Snippet: Fig. 3 | JUNB-HDAC1 complex represses inﬂammatory signals and cJUN. a Heatmap showing expression of cytokines present in the core enrichment of the gene sets shown in Supplementary Fig. 3a–d, for JUNB silencing (siJUNB; n = 3 biological replicates) versus control siRNA (siCtrl; n = 2 biological replicates) in CAPAN1 cells. Cell color indicates z score. b qRT-PCR analysis for indicated target genes in siJUNB conditions (red), normalized to siCtrl (gray), in CAPAN1. Relative mRNA expression with mean ± s.d. shown. n = 3 biological replicates. Two-tailed Student’s t-test with Welch’s correction. Coverage of JUNB ChIP-seq data in CAPAN121, as well as publicly available H3K27ac25 data, for loci of cJUN (c), IL1A/B (d), and CXCL9/10/11 (e). ChIP-qPCR validation regions are indicated. f Gene set enrichment analysis for curated signatures (C2) of the Molecular Signature Data- base (MSigDB) for siJUNB versus siCtrl in CAPAN1 cells. Normalized enrichment score (NES) and FDR q value are indicated. Immunoblot for JUNB, HDAC1, and β- actin after JUNB pulldown, IgG isotype control or input in CAPAN1 (g) and CAPAN2 (h). n = 3 biological replicates. i, j, ChIP-qPCR for regions indicated in (c–e), showing signal relative to input for JUNB (i) and HDAC1 (j) pulldown with

Article Snippet: Established human PDAC cell lines CAPAN1, CAPAN2 and MiaPaCa2 were purchased from ATCC (Manassas, VA) and authenticated (CAPAN1, RRID:CVCL_0237), (CAPAN2, RRID:CVCL_0026), (MiaPaCa2:CVCL_0428) by the German Collection ofMicroorganisms and cell culture GmbH (DSMZ).

Techniques: Expressing, Control, Quantitative RT-PCR, Two Tailed Test, ChIP-sequencing, ChIP-qPCR, Biomarker Discovery, Western Blot

Fig. 6 | Spatial TNF-α expression promotes macrophage inﬁltration and T-cell exclusion. a–h IHC analysis in 105 PDAC patients of the Princess Margaret Cancer Centre (PMCC) for TNF-α expression. a Spatial heterogeneity of TNF-α expression within different TMA cores of each patient. b IHC for TNF-α and CD68 in cores classiﬁed as TNF-αlow and TNF-αhigh. Scale bar 200 μm. c, d Quantiﬁcation of (b), in TNF-αlow, TNF-αintermediate (TNF-αint), and TNF-αhigh expression per patient (c) and per TMA core across all patients (d). c TNF-αhigh, n = 30 patients; TNF-αint, n = 28 patients; TNF-αlow, n = 32 patients. d TNF-αhigh, n = 87 cores; TNF-αint, n = 92 cores; TNF-αlow, n = 96 cores. e Lymphoid compartment distribution in TNF-αlow/int/high

Journal: Nature Communications

Article Title: Spatial tumor immune heterogeneity facilitates subtype co-existence and therapy response in pancreatic cancer

doi: 10.1038/s41467-024-55330-7

Figure Lengend Snippet: Fig. 6 | Spatial TNF-α expression promotes macrophage inﬁltration and T-cell exclusion. a–h IHC analysis in 105 PDAC patients of the Princess Margaret Cancer Centre (PMCC) for TNF-α expression. a Spatial heterogeneity of TNF-α expression within different TMA cores of each patient. b IHC for TNF-α and CD68 in cores classiﬁed as TNF-αlow and TNF-αhigh. Scale bar 200 μm. c, d Quantiﬁcation of (b), in TNF-αlow, TNF-αintermediate (TNF-αint), and TNF-αhigh expression per patient (c) and per TMA core across all patients (d). c TNF-αhigh, n = 30 patients; TNF-αint, n = 28 patients; TNF-αlow, n = 32 patients. d TNF-αhigh, n = 87 cores; TNF-αint, n = 92 cores; TNF-αlow, n = 96 cores. e Lymphoid compartment distribution in TNF-αlow/int/high

Techniques: Expressing

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