u937 monocytic cells  (ATCC)

Journal: Frontiers in Immunology

Article Title: Transcriptomic characterization of platelet-rich fibrin-induced macrophage responses identifies U937 cells as a sensitive bioassay

doi: 10.3389/fimmu.2026.1722342

Figure Lengend Snippet: PCA,principal component analysis of differentially expressed genes in PRF-activated U937 and THP-1 macrophages. The plot depicts the sample distribution in a two-dimensional space defined by the first and second principal components of the covariance matrix. Gene expression values were normalized and expressed as logCPM, log counts per million.

Article Snippet: The human monocytic cell lines U937 (CRL-1593.2) and THP-1 (TIB-202) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Gene Expression

Journal: Frontiers in Immunology

Article Title: Transcriptomic characterization of platelet-rich fibrin-induced macrophage responses identifies U937 cells as a sensitive bioassay

doi: 10.3389/fimmu.2026.1722342

Figure Lengend Snippet: Volcano plot analysis of differentially expressed genes in U937 and THP-1 macrophages treated with PRF. Volcano plots illustrate upregulated (red) and downregulated (blue) genes in U937 and THP-1 cells following PRF exposure. The annotated data points represent the 50 genes with the greatest Euclidean (Manhattan) distance from the origin that exceed the defined significance thresholds (dashed lines). Thresholds were set at –log 10 (p) ≥ 2.0 and |log 2 fold change| ≥ 3.0.

Article Snippet: The human monocytic cell lines U937 (CRL-1593.2) and THP-1 (TIB-202) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques:

Journal: Frontiers in Immunology

Article Title: Transcriptomic characterization of platelet-rich fibrin-induced macrophage responses identifies U937 cells as a sensitive bioassay

doi: 10.3389/fimmu.2026.1722342

Figure Lengend Snippet: Heatmap of differentially expressed genes in U937 and THP-1 macrophages exposed to PRF lysates. The heatmap visualizes the 395 upregulated and 372 downregulated genes in U937 cells, and the 116 upregulated and 149 downregulated genes in THP-1 cells, following exposure to PRF lysates. Differentially expressed genes were defined by an adjusted p-value < 0.05. The dataset integrates transcriptional data from three independent PRF preparations. Color gradients indicate relative expression levels, with red representing upregulation and blue representing downregulation; color intensity reflects the magnitude of the expression change.

Article Snippet: The human monocytic cell lines U937 (CRL-1593.2) and THP-1 (TIB-202) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: Transcriptomic characterization of platelet-rich fibrin-induced macrophage responses identifies U937 cells as a sensitive bioassay

doi: 10.3389/fimmu.2026.1722342

Figure Lengend Snippet: Venn analysis of differentially expressed genes in U937 and THP-1 macrophages treated with PRF lysates. Venn diagrams show the overlap of upregulated and downregulated genes between U937 and THP-1 cells after exposure to PRF lysates (adjusted p < 0.05). A total of 28 genes were commonly upregulated, and 20 genes were commonly downregulated in both cell lines.

Article Snippet: The human monocytic cell lines U937 (CRL-1593.2) and THP-1 (TIB-202) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques:

Journal: Frontiers in Immunology

Article Title: Transcriptomic characterization of platelet-rich fibrin-induced macrophage responses identifies U937 cells as a sensitive bioassay

doi: 10.3389/fimmu.2026.1722342

Figure Lengend Snippet: Functional enrichment analysis of upregulated genes in U937 macrophages stimulated with PRF lysates. Functional enrichment analysis, also known as over-representation analysis (ORA), was performed using the g:Profiler web-based platform. The plot depicts the top significantly enriched pathways derived from Gene Ontology and other integrated databases. Pathways are ranked and labeled numerically according to significance. P-values were adjusted for multiple testing using the Benjamini–Hochberg correction method (Padj).

Article Snippet: The human monocytic cell lines U937 (CRL-1593.2) and THP-1 (TIB-202) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Functional Assay, Derivative Assay, Labeling

Journal: Frontiers in Immunology

Article Title: Transcriptomic characterization of platelet-rich fibrin-induced macrophage responses identifies U937 cells as a sensitive bioassay

doi: 10.3389/fimmu.2026.1722342

Figure Lengend Snippet: Functional enrichment analysis of downregulated genes in U937 macrophages stimulated with PRF lysates. Functional enrichment analysis (over-representation analysis, ORA) was performed using the g:Profiler web-based platform. The figure displays the top significantly enriched pathways identified from Gene Ontology and other integrated databases, ranked and labeled numerically by significance. p-values were adjusted for multiple testing using the Benjamini–Hochberg correction method (Padj).

Article Snippet: The human monocytic cell lines U937 (CRL-1593.2) and THP-1 (TIB-202) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Functional Assay, Labeling

Journal: Frontiers in Immunology

Article Title: Transcriptomic characterization of platelet-rich fibrin-induced macrophage responses identifies U937 cells as a sensitive bioassay

doi: 10.3389/fimmu.2026.1722342

Figure Lengend Snippet: RT–qPCR validation of selected genes in U937 macrophages stimulated with PRF lysates. U937 macrophages were exposed to PRF lysates for 48 hours before quantitative RT–PCR analysis. Data represent the median of four independent experiments and two gene panels (A, B) .

Article Snippet: The human monocytic cell lines U937 (CRL-1593.2) and THP-1 (TIB-202) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Quantitative RT-PCR, Biomarker Discovery

Journal: Frontiers in Immunology

Article Title: Transcriptomic characterization of platelet-rich fibrin-induced macrophage responses identifies U937 cells as a sensitive bioassay

doi: 10.3389/fimmu.2026.1722342

Figure Lengend Snippet: RT–qPCR and ELISA analysis of inflammatory gene expression in U937 and THP-1 macrophages stimulated with PRF lysates. U937 (A, B) and THP-1 (C, D) macrophages were exposed to PRF lysates for 48 hours before quantitative RT–PCR analysis. ELISA quantified CXCL8 protein levels in the corresponding cell culture supernatants. Data represent the median of independent experiments.

Article Snippet: The human monocytic cell lines U937 (CRL-1593.2) and THP-1 (TIB-202) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Gene Expression, Cell Culture

Journal: bioRxiv

Article Title: TIM3 + Tumor Associated M2 Macrophages Impair Antitumor T Cell Immunity and Promote Gastric Cancer Progression and Peritoneal Metastasis

doi: 10.64898/2026.04.15.716939

Figure Lengend Snippet: A. Violin plot showing TIM3 expression across all cell types from scRNA-seq analysis of unfractionated live cells from 20 GCPM samples. B . Multi-parameter flow cytometric analysis of TIM3 and its ligands Gal-9 and CEACAM1 across different cell types in 54 GCPM samples (see Supplemental Table 1). C . CyTOF analysis of TIM3, Gal-9, and immune markers in six GCPM samples, using cytokeratin, CD45, CD34, and CD68 to define epithelial and immune populations, as described in Methods. D . Strong positive correlations between TIM3 and CD163 (R = 0.94, P = 1.4 × 10□¹□) and between TIM3 and Gal-9 (R = 0.87, P = 1 × 10□¹²), validated by scRNA-seq analysis. E . Positive correlations between TIM3 and M2 macrophage markers CD163 and CSF1R in GC tissues, analyzed using the GEPIA database. F . Representative co-immunofluorescence staining of TIM3 and CD163 in five GCPM samples. Scale bar, 20 μm. G . Schematic illustrating macrophage polarization from U937 monocytes to an M1 state followed by repolarization to an M2 state using GA0518 tumor cell–conditioned medium. H . qRT-PCR analysis of TIM3, Gal-9, and M2-associated markers (CCL2 and IL10) in U937-derived M1 and M2 macrophages. P values are indicated.

Article Snippet: Human monocyte U937 and murine macrophage RAW264.7 were purchased from ATCC, and both monocytes and macrophages were cultured in 7% FBS-RPMI.

Techniques: Expressing, Immunofluorescence, Staining, Quantitative RT-PCR, Derivative Assay

Journal: bioRxiv

Article Title: TIM3 + Tumor Associated M2 Macrophages Impair Antitumor T Cell Immunity and Promote Gastric Cancer Progression and Peritoneal Metastasis

doi: 10.64898/2026.04.15.716939

Figure Lengend Snippet: A. Schematic of the in vitro tumor cell invasion assay toward M2 macrophages with or without TIM3 knockout (KO). B. Effects of TIM3 KO in human U937 macrophages on GA0518 tumor cell invasion. Left: Western blot validation of TIM3 KO. Middle: representative invasion images. Right: quantification of invaded cells. C. Effects of TIM3 KO in murine RAW264.7 macrophages on KP-Luc2 tumor cell invasion, with validation, representative images, and quantification. . D-F . Co-inoculation of KP-Luc2 tumor cells with RAW264.7-derived M2 macrophages expressing TIM3 or TIM3 KO. ( D ) Representative tumor images. ( E ) Tumor growth curves and volumes. ( F ) Tumor weights at the experimental endpoint.

Article Snippet: Human monocyte U937 and murine macrophage RAW264.7 were purchased from ATCC, and both monocytes and macrophages were cultured in 7% FBS-RPMI.

Techniques: In Vitro, Invasion Assay, Knock-Out, Western Blot, Biomarker Discovery, Derivative Assay, Expressing

Journal: bioRxiv

Article Title: TIM3 + Tumor Associated M2 Macrophages Impair Antitumor T Cell Immunity and Promote Gastric Cancer Progression and Peritoneal Metastasis

doi: 10.64898/2026.04.15.716939

Figure Lengend Snippet: A. Schematic of in vitro co-culture of human or murine macrophages (control vs TIM3 KO) with PBMCs. B , Flow cytometry analysis of CD3□ or CD8□ T cells and intracellular cytokines, interferon-γ (IFNγ) and perforin in PBMCs after co-cultured with U937-TIM3 KO cells compared with U937-Control (Ctrl) cells. C , Flow cytometry of CD3□ or CD8□ T cells and intracellular IFNγ and perforin in PBMC co-cultured with murine RAW-TIM3KO macrophages versus RAW-Control. D. co-culturing of murine PBMCs with mouse bone-marrow-derived macrophages (mBM-Mø) under tumor cell derived conditioned medium (cGM) vs growth medium (GM), followed by anti-TIM3 antibody treatment. E . Flow cytometric quantification of CD3□/CD8□T cells and IFNγ production in CD45□ immune cells from malignant ascites co-cultured with U937 macrophages treated with anti-TIM3 antibody. F-G. Flow cytometric analysis of CD3 /CD8 T cell numbers (F) and IFNγ production (G) in tumors from the KP-Luc2 syngeneic model co-inoculated with RAW264.7 macrophages.

Article Snippet: Human monocyte U937 and murine macrophage RAW264.7 were purchased from ATCC, and both monocytes and macrophages were cultured in 7% FBS-RPMI.

Techniques: In Vitro, Co-Culture Assay, Control, Flow Cytometry, Cell Culture, Derivative Assay

Journal: bioRxiv

Article Title: TIM3 + Tumor Associated M2 Macrophages Impair Antitumor T Cell Immunity and Promote Gastric Cancer Progression and Peritoneal Metastasis

doi: 10.64898/2026.04.15.716939

Figure Lengend Snippet: A. Immunofluorescence staining of TIM3 in U937-derived M1 and M2 macrophages with or without TIM3 KO. B. Western blot confirmation of TIM3 KO in U937 macrophages. C. qRT-PCR analysis of CCL2 and IL10 expression in U937 M2 macrophages with or without TIM3 KO. D. qRT-PCR analysis of murine M2 macrophage markers (CCL2, Arg1, and IL10) in RAW264.7 macrophages with or without TIM3 KO. E , Cytokine array profiling of secreted factors from U937 TIM3 KO versus control cells using the RayBio C-Series Human Cytokine Antibody Array. F , Validation of seven selected cytokines/chemokines by qRT-PCR. P values are indicated.

Article Snippet: Human monocyte U937 and murine macrophage RAW264.7 were purchased from ATCC, and both monocytes and macrophages were cultured in 7% FBS-RPMI.

Techniques: Immunofluorescence, Staining, Derivative Assay, Western Blot, Quantitative RT-PCR, Expressing, Control, Ab Array, Biomarker Discovery

Journal: bioRxiv

Article Title: TIM3 + Tumor Associated M2 Macrophages Impair Antitumor T Cell Immunity and Promote Gastric Cancer Progression and Peritoneal Metastasis

doi: 10.64898/2026.04.15.716939

Figure Lengend Snippet: A. Violin plots showing CCL20, CCL4, CCL7, and IL1A expression across cell types in 20 GCPM samples by scRNA-seq; B. Relative expression of CCL20, CCL1, CCL4, and CCL7 in GC tumor versus normal tissues from the GSE33335 cohort. C. CCL20 expression in U937-derived M1 and M2 macrophages assessed by qRT-PCR and ELISA. D. ELISA measurement of secreted CCL20 from human U937 and murine RAW264.7 macrophages with or without TIM3 KO. E , Flow cytometric analysis of IFNγ and perforin production in CD3 and CD8 T cells co-cultured with U937 macrophages in the presence of recombinant CCL20. F. Tumor cell invasion toward U937 control or TIM3 KO macrophages in the presence of recombinant CCL20. G . Phospho-kinase array analysis comparing U937 TIM3 KO and control macrophages after M2 repolarization. H , Western blot validation of TIM3 and selected kinases in U937 macrophages. I. Effect of GDC-0994 on phosphorylation of p90RSK1/2 was determined by Western blot analysis. J. qRT-PCR analysis of CCL20 expression following GDC-0994 treatment compared with TIM3 KO.

Article Snippet: Human monocyte U937 and murine macrophage RAW264.7 were purchased from ATCC, and both monocytes and macrophages were cultured in 7% FBS-RPMI.

Techniques: Expressing, Derivative Assay, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Cell Culture, Recombinant, Control, Western Blot, Biomarker Discovery, Phospho-proteomics