human microglia cell line hmc3  (Elabscience Biotechnology)

Journal: Scientific Reports

Article Title: Interferon-β and FTY720 ameliorate progressive CNS inflammation via SOCS1-associated astrocyte signaling

doi: 10.1038/s41598-026-45303-9

Figure Lengend Snippet: Glial cells respond to FTY720 and IFN-β treatment with cell type–specific modulation of inflammatory and regenerative programs. Primary cultures of murine glial cells were treated with vehicle, FTY720, or the combination of FTY720 and IFN-β overnight and then stimulated with IL-1β and TNF-ɑ for 4 h. qPCR of the indicated genes was performed in ( A-D ) microglia and ( n = 3 biological replicates) and ( E–F ) astrocytes (n = 6 biological replicates). ( G ) Astrocyte-conditioned medium (ACM) was generated as described in Fig. H. In brief, primary astrocytes were activated with IL-1β (10 ng/mL) and TNF-α (5 ng/mL) for 24 h, followed by treatment with FTY720/IFN-β and either the control peptide or pJAK2(1001–1013) (20 µM) for 6 h in FCS-free medium. The medium was replaced the next day and collected after an additional 24 h. The harvested ACM was added to activated astrocytes under the respective conditions for subsequent qPCR analysis. qPCR was performed of indicated genes (n = 6 biological replicates). ( H ) Migration assay was performed as described in Fig. H, and the number of migrated monocytes was measured after 3 h ( n = 5 biological replicates). ( I ) Activated human microglia (HMC3 cells; n = 6) and ( J ) human astrocytes (Human astrocytes Catalog #1800, ScienCell; n = 3) were treated with vehicle, FTY720 and vehicle, or the combination of FTY720 and IFN-β overnight and then stimulated with IL-1β and TNF-ɑ for 4 h. qPCR of the indicated genes was performed. All qPCR results are displayed by fold change in mRNA expression. P-values of p < 0.05 were considered significant and determined by one-way ANOVA followed by Tukey´s multiple-comparisons test.

Article Snippet: For further in vitro experiments we used the human microglia cell line HMC3 (American Type Culture Collection [ATCC], Manassas, VA, USA; CRL-3304) and primary human astrocytes (ScienCell Research Laboratories, Carlsbad, CA, USA; Human Astrocytes, Catalog #1800).

Techniques: Generated, Control, Migration, Expressing

Journal: Nature Communications

Article Title: A common 19 bp APOE enhancer deletion is protective against Alzheimer’s disease in African Americans

doi: 10.1038/s41467-026-68808-3

Figure Lengend Snippet: a Schematic of the luciferase experimental design and constructs. The APOE 3′UTR ( APOE 3′UTR only) plus ~400 bp of downstream DNA containing the 19 bp deletion region and SPI1 binding site were cloned into a psiCheck2.2 dual luciferase reporter construct. b Renilla:Firefly luciferase expression data in HMC3 cells after delivery of APOE SPI1 WT sequence ( p = 0.0016 relative to the APOE 3′UTR only) or delivery of the 19 bp deletion [ p = 0.8252; one-way ANOVA, n = 12 samples/group (3 biological replicates, 4 technical replicates), ± SD]. c –e HMC3 gene expression following SPI1 overexpression. c Effect of SPI1 overexpression on c APOE expression ( p = 0.3831, F = 1.789, unpaired two-tailed t-test, n = 12 control and 12 SPI1 biological replicates derived from the average of 4 technical replicates/sample, ± SEM), d APOC1 expression ( p = <0.0001, F = 2.987, unpaired two-tailed t-test, n = 11 control and 12 SPI1 biological replicates derived from the average of 4 technical replicates/sample, ± SEM), and e lncRNA ENSG00000280087 expression ( p = 0.0170, F = 5.414, unpaired two-tailed t-test, n = 12 control and 10 SPI1 biological replicates derived from the average of 4 technical replicates/sample, ± SEM). f – h HMC3 gene expression following Aβ 1:42 treatment. f Effect of Aβ 1:42 treatment on APOE expression ( p = 0.2098, F = 1.243, unpaired two-tailed t-test, n = 12 control and 12 Aβ biological replicates derived from the average of 4 technical replicates/sample, ± SEM), g APOC1 expression ( p = 0.0015, F = 4.657, unpaired two-tailed t-test, n = 11 control and 12 Aβ biological replicates derived from the average of 4 technical replicates/sample, ± SEM) and h, lncRNA ENSG00000280087 expression ( p = 0.0255, F = 2.502, unpaired two-tailed t-test, n = 12 control and 12 Aβ biological replicates derived from the average of 4 technical replicates/sample, ± SEM) in HMC3 cells.

Article Snippet: HMC3 human microglia cell lines (ATCC CRL-3304) were grown in EMEM media and seeded at 3.34 × 10 4 cells plated in 12-well plates for luciferase assays.

Techniques: Luciferase, Construct, Binding Assay, Clone Assay, Expressing, Sequencing, Gene Expression, Over Expression, Two Tailed Test, Control, Derivative Assay

Journal: bioRxiv

Article Title: Modelling ferroptosis in a human microglial line by sequential exposure to iron and GPX4 inhibition

doi: 10.64898/2026.01.19.700282

Figure Lengend Snippet: A) Dose-response curves with RSL3 treatment (0-1 µM) for 2 hr with measurement of (i) levels of lipid peroxidation (BODIPY 581/591 C11) and (ii) cell viability (DRAQ7). Dotted line represents control values from DMSO-treated samples against which other samples are compared for statistical significance. N=3, one-way ANOVA with Šidák’s test, *p < 0.05, ***p < 0.001. B) Cytofluorimetric measurement of (i) levels of lipid peroxidation and of (ii) cell viability, following treatment with Erastin (10 µM), RSL3 (200 nM) or DMSO (control). N=3, one-way ANOVA with Tukey’s test, **p < 0.005, ***p < 0.001. C) (i) Overview of the experimental design. Dose-response curves of iron-loading (0-500 µM) for 24 hr, followed by treatment with DMSO (circles) or RSL3 (200 nM, triangles) for 2 hr measuring levels of (ii) lipid peroxidation and (iii) cell viability. Dotted lines represent control values from samples without iron against which the other samples are compared for statistical significance. N=3, one-way ANOVA with Dunett’s test, *p < 0.05. D) Rescue phenotype with Fer-1 (10 µM) after single or combined treatments of iron (50 µM) and RSL3 (200 nM) measuring levels of (i) lipid peroxidation and (i) cell viability. N=3, one-way ANOVA with Tukey’s test, *p < 0.05, ***p < 0.001. E) Representative images of HMC3 cells stained with (i) CC3 antibody (Green). Menadione treatment was used as positive control to induce apoptosis. (ii) Quantification of CC3 positive nuclei, plotted as percentage of total number of nuclei defined with Hoechst. Scale bar represents 100 µm. (iii) HMC3 survival measuring total nuclei (Hoechst) and plotted as percentage over DMSO-treated control. N=3, one-way ANOVA with Tukey’s test, **p < 0.01.

Article Snippet: Human microglia HMC3 cell line was purchased from ATCC (CRL-3304) and cultured in MEM containing Earl’s Salts and glutamine (Gibco, 31095-029) supplemented with 10% FBS (Gibco, 16140-063), 1% penicillin and streptomycin (Gibco, 15140-122) at 37°C and in a humidified atmosphere containing 5% CO2.

Techniques: Control, Staining, Positive Control

Journal: bioRxiv

Article Title: Modelling ferroptosis in a human microglial line by sequential exposure to iron and GPX4 inhibition

doi: 10.64898/2026.01.19.700282

Figure Lengend Snippet: A) (i) Representative images of HMC3 cells stained with CellMask Green and CellRox Orange. The staining was performed after 24 hr iron-loading of the cells (50 µM FAC) followed by treatment for 2 hr with DMSO or RSL3 (200 nM) +/- Fer-1 (10 µM). Scale bar=100µm. (ii) Quantification of CellRox intensity for DMSO (grey) or RSL3 (orange). N=3, one-way ANOVA with Šidák’s test, ***p < 0.001. B) MitoSOX labelling and quantification using flow cytometry to detect (i) mitochondrial superoxide and (ii) total ROS. N=4, one-way ANOVA with Šidák’s test, **p < 0.00. C) Workflow of cell culture for the model and cell painting analysis. Scale bar=100µm. D) Contingency tables listing the numbers of most important features per organelle/compartment in the top 35-40 features, ranked by Random Forest classifier, to differentiate treatment conditions. Cell summaries were excluded from Fisher Exact Test as too few of these features were present.

Article Snippet: Human microglia HMC3 cell line was purchased from ATCC (CRL-3304) and cultured in MEM containing Earl’s Salts and glutamine (Gibco, 31095-029) supplemented with 10% FBS (Gibco, 16140-063), 1% penicillin and streptomycin (Gibco, 15140-122) at 37°C and in a humidified atmosphere containing 5% CO2.

Techniques: Staining, Flow Cytometry, Cell Culture

Journal: bioRxiv

Article Title: Modelling ferroptosis in a human microglial line by sequential exposure to iron and GPX4 inhibition

doi: 10.64898/2026.01.19.700282

Figure Lengend Snippet: A) Overview of sample collection process. B) Differentially abundant lipids represented on Volcano plots for all the treatments. The top 10 significant lipids, ranked by adj. p-value and log2 fold-change are highlighted alongside most significant sterols. C) Heatmap showing abundance of lipids averaged across replicates and LipidMaps classes. Lipid abundance data is on a unit-variance scale to make analytes comparable with each other. D) Lipid enrichment analysis barplot showing top 10 lipid classes upregulated in ferroptotic HMC3 (positive log2 fold change of IronRSL3 vs DMSO). Sub-classes obtained from LipidMaps. MWU test to identify differential lipids (p <0.05) and ORA performed on lipid classes). E) Barplots showing top 20 (i) LipidMaps classes and (ii) lipid species restored by Fer-1 treatment (negative log2 fold change of IronRSL3-Fer1 vs IronRSL3).

Article Snippet: Human microglia HMC3 cell line was purchased from ATCC (CRL-3304) and cultured in MEM containing Earl’s Salts and glutamine (Gibco, 31095-029) supplemented with 10% FBS (Gibco, 16140-063), 1% penicillin and streptomycin (Gibco, 15140-122) at 37°C and in a humidified atmosphere containing 5% CO2.

Techniques: