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Magnetic Bead Sorting, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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magnetic beads cell sorting  (Miltenyi Biotec)
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Coculture with Non-small <t>Cell</t> Lung Cancer (NSCLC) cells lead to profound transcriptomic endothelial cell alterations. A ) Schematic of the 2D in vitro coculture system where human endothelial cells (ECs; HUVECs) were cocultured with/without NSCLC cell lines during 48h. Thereafter, normal (NECs) and NSCLC-cocultured ECs (NSCLC-TECs) were isolated using anti-CD31 antibody-coated <t>magnetic</t> <t>beads,</t> and 3’RNA-seq, kinome and functional validations were performed. B ) Immunostaining for EC-specific markers (vWF in red and CD31 in green) revealed the spatial organization of HUVECs in cocultures. Scale bar: 50 μm. C ) Flow cytometric analysis of different cocultured cells stained for CD31 before (i) and after (ii) CD31-enrichment. D ) Principal component analysis considering the top 100 highly-variable genes. Samples segregate into 3 major groups with H1755-TECs being more similar to NECs as compared to the other NSCLC-TECs. n =8 per group. E ) Correlation heatmap of the top 350 highly variable genes in NECs/NSCLC-TECs. F ) Gene set enrichment analysis in all NSCLC-TECs versus NECs. Pathways enriched or downregulated in NSCLC-TECs appear in red and blue, respectively. Adjusted p -value < 0.05. G - H ) Meta-analysis between NSCLC-TECs and HUVECs cocultured similarly with the breast cancer MDA-MB-231 cell line. Results show congruent up/downregulated targets at the G ) gene and H ) gene set levels
Magnetic Beads Cell Sorting, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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magnetic bead based cell sorting  (Miltenyi Biotec)
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Coculture with Non-small <t>Cell</t> Lung Cancer (NSCLC) cells lead to profound transcriptomic endothelial cell alterations. A ) Schematic of the 2D in vitro coculture system where human endothelial cells (ECs; HUVECs) were cocultured with/without NSCLC cell lines during 48h. Thereafter, normal (NECs) and NSCLC-cocultured ECs (NSCLC-TECs) were isolated using anti-CD31 antibody-coated <t>magnetic</t> <t>beads,</t> and 3’RNA-seq, kinome and functional validations were performed. B ) Immunostaining for EC-specific markers (vWF in red and CD31 in green) revealed the spatial organization of HUVECs in cocultures. Scale bar: 50 μm. C ) Flow cytometric analysis of different cocultured cells stained for CD31 before (i) and after (ii) CD31-enrichment. D ) Principal component analysis considering the top 100 highly-variable genes. Samples segregate into 3 major groups with H1755-TECs being more similar to NECs as compared to the other NSCLC-TECs. n =8 per group. E ) Correlation heatmap of the top 350 highly variable genes in NECs/NSCLC-TECs. F ) Gene set enrichment analysis in all NSCLC-TECs versus NECs. Pathways enriched or downregulated in NSCLC-TECs appear in red and blue, respectively. Adjusted p -value < 0.05. G - H ) Meta-analysis between NSCLC-TECs and HUVECs cocultured similarly with the breast cancer MDA-MB-231 cell line. Results show congruent up/downregulated targets at the G ) gene and H ) gene set levels
Magnetic Bead Based Cell Sorting, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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cd45 magnetic bead sorting  (Miltenyi Biotec)
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Coculture with Non-small <t>Cell</t> Lung Cancer (NSCLC) cells lead to profound transcriptomic endothelial cell alterations. A ) Schematic of the 2D in vitro coculture system where human endothelial cells (ECs; HUVECs) were cocultured with/without NSCLC cell lines during 48h. Thereafter, normal (NECs) and NSCLC-cocultured ECs (NSCLC-TECs) were isolated using anti-CD31 antibody-coated <t>magnetic</t> <t>beads,</t> and 3’RNA-seq, kinome and functional validations were performed. B ) Immunostaining for EC-specific markers (vWF in red and CD31 in green) revealed the spatial organization of HUVECs in cocultures. Scale bar: 50 μm. C ) Flow cytometric analysis of different cocultured cells stained for CD31 before (i) and after (ii) CD31-enrichment. D ) Principal component analysis considering the top 100 highly-variable genes. Samples segregate into 3 major groups with H1755-TECs being more similar to NECs as compared to the other NSCLC-TECs. n =8 per group. E ) Correlation heatmap of the top 350 highly variable genes in NECs/NSCLC-TECs. F ) Gene set enrichment analysis in all NSCLC-TECs versus NECs. Pathways enriched or downregulated in NSCLC-TECs appear in red and blue, respectively. Adjusted p -value < 0.05. G - H ) Meta-analysis between NSCLC-TECs and HUVECs cocultured similarly with the breast cancer MDA-MB-231 cell line. Results show congruent up/downregulated targets at the G ) gene and H ) gene set levels
Cd45 Magnetic Bead Sorting, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti mouse igg magnetic bead magnetic activated cell sorting macs  (Miltenyi Biotec)
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Coculture with Non-small <t>Cell</t> Lung Cancer (NSCLC) cells lead to profound transcriptomic endothelial cell alterations. A ) Schematic of the 2D in vitro coculture system where human endothelial cells (ECs; HUVECs) were cocultured with/without NSCLC cell lines during 48h. Thereafter, normal (NECs) and NSCLC-cocultured ECs (NSCLC-TECs) were isolated using anti-CD31 antibody-coated <t>magnetic</t> <t>beads,</t> and 3’RNA-seq, kinome and functional validations were performed. B ) Immunostaining for EC-specific markers (vWF in red and CD31 in green) revealed the spatial organization of HUVECs in cocultures. Scale bar: 50 μm. C ) Flow cytometric analysis of different cocultured cells stained for CD31 before (i) and after (ii) CD31-enrichment. D ) Principal component analysis considering the top 100 highly-variable genes. Samples segregate into 3 major groups with H1755-TECs being more similar to NECs as compared to the other NSCLC-TECs. n =8 per group. E ) Correlation heatmap of the top 350 highly variable genes in NECs/NSCLC-TECs. F ) Gene set enrichment analysis in all NSCLC-TECs versus NECs. Pathways enriched or downregulated in NSCLC-TECs appear in red and blue, respectively. Adjusted p -value < 0.05. G - H ) Meta-analysis between NSCLC-TECs and HUVECs cocultured similarly with the breast cancer MDA-MB-231 cell line. Results show congruent up/downregulated targets at the G ) gene and H ) gene set levels
Anti Mouse Igg Magnetic Bead Magnetic Activated Cell Sorting Macs, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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cell sorting magnetic bead cd56  (Miltenyi Biotec)
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Coculture with Non-small <t>Cell</t> Lung Cancer (NSCLC) cells lead to profound transcriptomic endothelial cell alterations. A ) Schematic of the 2D in vitro coculture system where human endothelial cells (ECs; HUVECs) were cocultured with/without NSCLC cell lines during 48h. Thereafter, normal (NECs) and NSCLC-cocultured ECs (NSCLC-TECs) were isolated using anti-CD31 antibody-coated <t>magnetic</t> <t>beads,</t> and 3’RNA-seq, kinome and functional validations were performed. B ) Immunostaining for EC-specific markers (vWF in red and CD31 in green) revealed the spatial organization of HUVECs in cocultures. Scale bar: 50 μm. C ) Flow cytometric analysis of different cocultured cells stained for CD31 before (i) and after (ii) CD31-enrichment. D ) Principal component analysis considering the top 100 highly-variable genes. Samples segregate into 3 major groups with H1755-TECs being more similar to NECs as compared to the other NSCLC-TECs. n =8 per group. E ) Correlation heatmap of the top 350 highly variable genes in NECs/NSCLC-TECs. F ) Gene set enrichment analysis in all NSCLC-TECs versus NECs. Pathways enriched or downregulated in NSCLC-TECs appear in red and blue, respectively. Adjusted p -value < 0.05. G - H ) Meta-analysis between NSCLC-TECs and HUVECs cocultured similarly with the breast cancer MDA-MB-231 cell line. Results show congruent up/downregulated targets at the G ) gene and H ) gene set levels
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Coculture with Non-small Cell Lung Cancer (NSCLC) cells lead to profound transcriptomic endothelial cell alterations. A ) Schematic of the 2D in vitro coculture system where human endothelial cells (ECs; HUVECs) were cocultured with/without NSCLC cell lines during 48h. Thereafter, normal (NECs) and NSCLC-cocultured ECs (NSCLC-TECs) were isolated using anti-CD31 antibody-coated magnetic beads, and 3’RNA-seq, kinome and functional validations were performed. B ) Immunostaining for EC-specific markers (vWF in red and CD31 in green) revealed the spatial organization of HUVECs in cocultures. Scale bar: 50 μm. C ) Flow cytometric analysis of different cocultured cells stained for CD31 before (i) and after (ii) CD31-enrichment. D ) Principal component analysis considering the top 100 highly-variable genes. Samples segregate into 3 major groups with H1755-TECs being more similar to NECs as compared to the other NSCLC-TECs. n =8 per group. E ) Correlation heatmap of the top 350 highly variable genes in NECs/NSCLC-TECs. F ) Gene set enrichment analysis in all NSCLC-TECs versus NECs. Pathways enriched or downregulated in NSCLC-TECs appear in red and blue, respectively. Adjusted p -value < 0.05. G - H ) Meta-analysis between NSCLC-TECs and HUVECs cocultured similarly with the breast cancer MDA-MB-231 cell line. Results show congruent up/downregulated targets at the G ) gene and H ) gene set levels

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: In vitro models to mimic tumor endothelial cell-mediated immune cell reprogramming in lung adenocarcinoma

doi: 10.1186/s13046-025-03576-4

Figure Lengend Snippet: Coculture with Non-small Cell Lung Cancer (NSCLC) cells lead to profound transcriptomic endothelial cell alterations. A ) Schematic of the 2D in vitro coculture system where human endothelial cells (ECs; HUVECs) were cocultured with/without NSCLC cell lines during 48h. Thereafter, normal (NECs) and NSCLC-cocultured ECs (NSCLC-TECs) were isolated using anti-CD31 antibody-coated magnetic beads, and 3’RNA-seq, kinome and functional validations were performed. B ) Immunostaining for EC-specific markers (vWF in red and CD31 in green) revealed the spatial organization of HUVECs in cocultures. Scale bar: 50 μm. C ) Flow cytometric analysis of different cocultured cells stained for CD31 before (i) and after (ii) CD31-enrichment. D ) Principal component analysis considering the top 100 highly-variable genes. Samples segregate into 3 major groups with H1755-TECs being more similar to NECs as compared to the other NSCLC-TECs. n =8 per group. E ) Correlation heatmap of the top 350 highly variable genes in NECs/NSCLC-TECs. F ) Gene set enrichment analysis in all NSCLC-TECs versus NECs. Pathways enriched or downregulated in NSCLC-TECs appear in red and blue, respectively. Adjusted p -value < 0.05. G - H ) Meta-analysis between NSCLC-TECs and HUVECs cocultured similarly with the breast cancer MDA-MB-231 cell line. Results show congruent up/downregulated targets at the G ) gene and H ) gene set levels

Article Snippet: After 48 h, monoculture (HUVECs alone) or coculture (HUVECs + A549/HUVECs + H1755/HUVECs + H1975) are detached with TrypLE Select (1X) (Gibco TM ,12563029) and HUVECs were positively selected for CD31 by magnetic beads cell sorting (MACS, Miltenyi Biotec, 130-091−935) using LS separation columns (Miltenyi Biotec, 130-042−401) following the manufacturer’s instructions.

Techniques: In Vitro, Isolation, Magnetic Beads, Functional Assay, Immunostaining, Staining