This content is restricted to AACP Members. Click here to become a member.
Abstract
Discussion Forum (0)
Poster Category: NIA
AACP Section: Other
Objectives: To develop a 3D organotypic model of T-cell recruitment to the omental tumor microenvironment, the primary metastatic niche of ovarian cancer. Further, to evaluate the impact of the type 2 diabetes medication metformin on T-cell transendothelial migration in this novel model system.
Methods: The prototype 3D model included T-cells (Jurkat), mesothelial cells (MeT5A), fibroblasts (HDFa) and endothelial cells (HuVEC). Mesothelial cells and fibroblasts were separated from T-cells by a transwell chamber containing endothelial cells and collagen matrix on the porous mesh. The co-culture was established systematically over 72h prior to addition of T-cells and endpoint assay. T-cells were allowed to invade for 2h to model transendothelial migration towards the fibroblast/mesothelial cell layer or separated from the 3D co-culture by transwell for 24h for phenotyping by flow cytometry and RNA sequencing. In evaluation of the activity of metformin, cells were pretreated separately for 72h and then maintained during co-culture of the complete 3D model (250μM metformin). RNA sequencing data was analyzed using DEseq2 and differentially expressed genes were evaluated by Gene Set Enrichment Analysis.
Results: Using this model system, mesothelial cells and fibroblasts promoted T-cell transendothelial migration similar to chemokine (SDF1a) treatment alone. Metformin pretreatment and maintenance enhanced the ability of T-cells to invade through the endothelial monolayer. Phenotypically, metformin exposure favored a shift to cytotoxic T-cells as indicated by an increased CD8+ population. Pathway analysis suggested significant increases in cytoskeletal rearrangement.
Conclusion: Optimization of the conditions for this 3D organotypic model may provide a framework for its use with primary cells derived from omental metastases. Preliminary findings using this model system have supported that metformin may enhance T-cell transendothelial migration into the omental TME.
AACP Section: Other
Objectives: To develop a 3D organotypic model of T-cell recruitment to the omental tumor microenvironment, the primary metastatic niche of ovarian cancer. Further, to evaluate the impact of the type 2 diabetes medication metformin on T-cell transendothelial migration in this novel model system.
Methods: The prototype 3D model included T-cells (Jurkat), mesothelial cells (MeT5A), fibroblasts (HDFa) and endothelial cells (HuVEC). Mesothelial cells and fibroblasts were separated from T-cells by a transwell chamber containing endothelial cells and collagen matrix on the porous mesh. The co-culture was established systematically over 72h prior to addition of T-cells and endpoint assay. T-cells were allowed to invade for 2h to model transendothelial migration towards the fibroblast/mesothelial cell layer or separated from the 3D co-culture by transwell for 24h for phenotyping by flow cytometry and RNA sequencing. In evaluation of the activity of metformin, cells were pretreated separately for 72h and then maintained during co-culture of the complete 3D model (250μM metformin). RNA sequencing data was analyzed using DEseq2 and differentially expressed genes were evaluated by Gene Set Enrichment Analysis.
Results: Using this model system, mesothelial cells and fibroblasts promoted T-cell transendothelial migration similar to chemokine (SDF1a) treatment alone. Metformin pretreatment and maintenance enhanced the ability of T-cells to invade through the endothelial monolayer. Phenotypically, metformin exposure favored a shift to cytotoxic T-cells as indicated by an increased CD8+ population. Pathway analysis suggested significant increases in cytoskeletal rearrangement.
Conclusion: Optimization of the conditions for this 3D organotypic model may provide a framework for its use with primary cells derived from omental metastases. Preliminary findings using this model system have supported that metformin may enhance T-cell transendothelial migration into the omental TME.
Poster Category: NIA
AACP Section: Other
Objectives: To develop a 3D organotypic model of T-cell recruitment to the omental tumor microenvironment, the primary metastatic niche of ovarian cancer. Further, to evaluate the impact of the type 2 diabetes medication metformin on T-cell transendothelial migration in this novel model system.
Methods: The prototype 3D model included T-cells (Jurkat), mesothelial cells (MeT5A), fibroblasts (HDFa) and endothelial cells (HuVEC). Mesothelial cells and fibroblasts were separated from T-cells by a transwell chamber containing endothelial cells and collagen matrix on the porous mesh. The co-culture was established systematically over 72h prior to addition of T-cells and endpoint assay. T-cells were allowed to invade for 2h to model transendothelial migration towards the fibroblast/mesothelial cell layer or separated from the 3D co-culture by transwell for 24h for phenotyping by flow cytometry and RNA sequencing. In evaluation of the activity of metformin, cells were pretreated separately for 72h and then maintained during co-culture of the complete 3D model (250μM metformin). RNA sequencing data was analyzed using DEseq2 and differentially expressed genes were evaluated by Gene Set Enrichment Analysis.
Results: Using this model system, mesothelial cells and fibroblasts promoted T-cell transendothelial migration similar to chemokine (SDF1a) treatment alone. Metformin pretreatment and maintenance enhanced the ability of T-cells to invade through the endothelial monolayer. Phenotypically, metformin exposure favored a shift to cytotoxic T-cells as indicated by an increased CD8+ population. Pathway analysis suggested significant increases in cytoskeletal rearrangement.
Conclusion: Optimization of the conditions for this 3D organotypic model may provide a framework for its use with primary cells derived from omental metastases. Preliminary findings using this model system have supported that metformin may enhance T-cell transendothelial migration into the omental TME.
AACP Section: Other
Objectives: To develop a 3D organotypic model of T-cell recruitment to the omental tumor microenvironment, the primary metastatic niche of ovarian cancer. Further, to evaluate the impact of the type 2 diabetes medication metformin on T-cell transendothelial migration in this novel model system.
Methods: The prototype 3D model included T-cells (Jurkat), mesothelial cells (MeT5A), fibroblasts (HDFa) and endothelial cells (HuVEC). Mesothelial cells and fibroblasts were separated from T-cells by a transwell chamber containing endothelial cells and collagen matrix on the porous mesh. The co-culture was established systematically over 72h prior to addition of T-cells and endpoint assay. T-cells were allowed to invade for 2h to model transendothelial migration towards the fibroblast/mesothelial cell layer or separated from the 3D co-culture by transwell for 24h for phenotyping by flow cytometry and RNA sequencing. In evaluation of the activity of metformin, cells were pretreated separately for 72h and then maintained during co-culture of the complete 3D model (250μM metformin). RNA sequencing data was analyzed using DEseq2 and differentially expressed genes were evaluated by Gene Set Enrichment Analysis.
Results: Using this model system, mesothelial cells and fibroblasts promoted T-cell transendothelial migration similar to chemokine (SDF1a) treatment alone. Metformin pretreatment and maintenance enhanced the ability of T-cells to invade through the endothelial monolayer. Phenotypically, metformin exposure favored a shift to cytotoxic T-cells as indicated by an increased CD8+ population. Pathway analysis suggested significant increases in cytoskeletal rearrangement.
Conclusion: Optimization of the conditions for this 3D organotypic model may provide a framework for its use with primary cells derived from omental metastases. Preliminary findings using this model system have supported that metformin may enhance T-cell transendothelial migration into the omental TME.
Metformin Promotes T-Cell Recruitment in a Novel 3D Model of Omental Metastasis
Silver Homa
Abstract
Discussion Forum (0)
Poster Category: NIA
AACP Section: Other
Objectives: To develop a 3D organotypic model of T-cell recruitment to the omental tumor microenvironment, the primary metastatic niche of ovarian cancer. Further, to evaluate the impact of the type 2 diabetes medication metformin on T-cell transendothelial migration in this novel model system.
Methods: The prototype 3D model included T-cells (Jurkat), mesothelial cells (MeT5A), fibroblasts (HDFa) and endothelial cells (HuVEC). Mesothelial cells and fibroblasts were separated from T-cells by a transwell chamber containing endothelial cells and collagen matrix on the porous mesh. The co-culture was established systematically over 72h prior to addition of T-cells and endpoint assay. T-cells were allowed to invade for 2h to model transendothelial migration towards the fibroblast/mesothelial cell layer or separated from the 3D co-culture by transwell for 24h for phenotyping by flow cytometry and RNA sequencing. In evaluation of the activity of metformin, cells were pretreated separately for 72h and then maintained during co-culture of the complete 3D model (250μM metformin). RNA sequencing data was analyzed using DEseq2 and differentially expressed genes were evaluated by Gene Set Enrichment Analysis.
Results: Using this model system, mesothelial cells and fibroblasts promoted T-cell transendothelial migration similar to chemokine (SDF1a) treatment alone. Metformin pretreatment and maintenance enhanced the ability of T-cells to invade through the endothelial monolayer. Phenotypically, metformin exposure favored a shift to cytotoxic T-cells as indicated by an increased CD8+ population. Pathway analysis suggested significant increases in cytoskeletal rearrangement.
Conclusion: Optimization of the conditions for this 3D organotypic model may provide a framework for its use with primary cells derived from omental metastases. Preliminary findings using this model system have supported that metformin may enhance T-cell transendothelial migration into the omental TME.
AACP Section: Other
Objectives: To develop a 3D organotypic model of T-cell recruitment to the omental tumor microenvironment, the primary metastatic niche of ovarian cancer. Further, to evaluate the impact of the type 2 diabetes medication metformin on T-cell transendothelial migration in this novel model system.
Methods: The prototype 3D model included T-cells (Jurkat), mesothelial cells (MeT5A), fibroblasts (HDFa) and endothelial cells (HuVEC). Mesothelial cells and fibroblasts were separated from T-cells by a transwell chamber containing endothelial cells and collagen matrix on the porous mesh. The co-culture was established systematically over 72h prior to addition of T-cells and endpoint assay. T-cells were allowed to invade for 2h to model transendothelial migration towards the fibroblast/mesothelial cell layer or separated from the 3D co-culture by transwell for 24h for phenotyping by flow cytometry and RNA sequencing. In evaluation of the activity of metformin, cells were pretreated separately for 72h and then maintained during co-culture of the complete 3D model (250μM metformin). RNA sequencing data was analyzed using DEseq2 and differentially expressed genes were evaluated by Gene Set Enrichment Analysis.
Results: Using this model system, mesothelial cells and fibroblasts promoted T-cell transendothelial migration similar to chemokine (SDF1a) treatment alone. Metformin pretreatment and maintenance enhanced the ability of T-cells to invade through the endothelial monolayer. Phenotypically, metformin exposure favored a shift to cytotoxic T-cells as indicated by an increased CD8+ population. Pathway analysis suggested significant increases in cytoskeletal rearrangement.
Conclusion: Optimization of the conditions for this 3D organotypic model may provide a framework for its use with primary cells derived from omental metastases. Preliminary findings using this model system have supported that metformin may enhance T-cell transendothelial migration into the omental TME.
Poster Category: NIA
AACP Section: Other
Objectives: To develop a 3D organotypic model of T-cell recruitment to the omental tumor microenvironment, the primary metastatic niche of ovarian cancer. Further, to evaluate the impact of the type 2 diabetes medication metformin on T-cell transendothelial migration in this novel model system.
Methods: The prototype 3D model included T-cells (Jurkat), mesothelial cells (MeT5A), fibroblasts (HDFa) and endothelial cells (HuVEC). Mesothelial cells and fibroblasts were separated from T-cells by a transwell chamber containing endothelial cells and collagen matrix on the porous mesh. The co-culture was established systematically over 72h prior to addition of T-cells and endpoint assay. T-cells were allowed to invade for 2h to model transendothelial migration towards the fibroblast/mesothelial cell layer or separated from the 3D co-culture by transwell for 24h for phenotyping by flow cytometry and RNA sequencing. In evaluation of the activity of metformin, cells were pretreated separately for 72h and then maintained during co-culture of the complete 3D model (250μM metformin). RNA sequencing data was analyzed using DEseq2 and differentially expressed genes were evaluated by Gene Set Enrichment Analysis.
Results: Using this model system, mesothelial cells and fibroblasts promoted T-cell transendothelial migration similar to chemokine (SDF1a) treatment alone. Metformin pretreatment and maintenance enhanced the ability of T-cells to invade through the endothelial monolayer. Phenotypically, metformin exposure favored a shift to cytotoxic T-cells as indicated by an increased CD8+ population. Pathway analysis suggested significant increases in cytoskeletal rearrangement.
Conclusion: Optimization of the conditions for this 3D organotypic model may provide a framework for its use with primary cells derived from omental metastases. Preliminary findings using this model system have supported that metformin may enhance T-cell transendothelial migration into the omental TME.
AACP Section: Other
Objectives: To develop a 3D organotypic model of T-cell recruitment to the omental tumor microenvironment, the primary metastatic niche of ovarian cancer. Further, to evaluate the impact of the type 2 diabetes medication metformin on T-cell transendothelial migration in this novel model system.
Methods: The prototype 3D model included T-cells (Jurkat), mesothelial cells (MeT5A), fibroblasts (HDFa) and endothelial cells (HuVEC). Mesothelial cells and fibroblasts were separated from T-cells by a transwell chamber containing endothelial cells and collagen matrix on the porous mesh. The co-culture was established systematically over 72h prior to addition of T-cells and endpoint assay. T-cells were allowed to invade for 2h to model transendothelial migration towards the fibroblast/mesothelial cell layer or separated from the 3D co-culture by transwell for 24h for phenotyping by flow cytometry and RNA sequencing. In evaluation of the activity of metformin, cells were pretreated separately for 72h and then maintained during co-culture of the complete 3D model (250μM metformin). RNA sequencing data was analyzed using DEseq2 and differentially expressed genes were evaluated by Gene Set Enrichment Analysis.
Results: Using this model system, mesothelial cells and fibroblasts promoted T-cell transendothelial migration similar to chemokine (SDF1a) treatment alone. Metformin pretreatment and maintenance enhanced the ability of T-cells to invade through the endothelial monolayer. Phenotypically, metformin exposure favored a shift to cytotoxic T-cells as indicated by an increased CD8+ population. Pathway analysis suggested significant increases in cytoskeletal rearrangement.
Conclusion: Optimization of the conditions for this 3D organotypic model may provide a framework for its use with primary cells derived from omental metastases. Preliminary findings using this model system have supported that metformin may enhance T-cell transendothelial migration into the omental TME.
{{ help_message }}
{{filter}}