Journal: iScience

Article Title: Spatial single-cell profiling identifies protein kinase Cδ-expressing microglia with anti-tumor function in glioblastoma

doi: 10.1016/j.isci.2025.114281

Figure Lengend Snippet: PKCδ in microglia contributes to the phagocytosis of BTICs (A) Schematic overview of the in vitro phagocytosis assay. (B and C) Representative IF images (B) and quantification (C) of phagocytosis of pHrodo-labeled S. aureus BioParticles by HMC3 microglia cells with PRKCD knockdown, in the presence or absence of niacin. (D) Phagocytosis assay using primary human microglia stimulated with niacin, with or without the PKC inhibitor CRT0066101. (E) Schematic of the in vitro phagocytosis assay using human BTICs labeled with pHrodo. (F and G) Representative images (F) and quantification (G) of phagocytosis of pHrodo-labeled human BTICs (BT012) by HMC3 cells with PRKCD knockdown. (H and I) Representative IF images (H) and quantification (I) of apoptotic BTICs (BT012 and BT025), determined by activated caspase-3/7 staining following co-culture with control or PRKCD -knockdown HMC3 cells, in the presence or absence of niacin. (J) Cell-type deconvolution of Visium spatial transcriptomics data from tumor-bearing mice treated with niacin. (K) Comparison of Prkcd expression between niacin-treated and control mice in spatial transcriptomics. (L) Quantification of Prkcd expression across spatial clusters. (M) IF staining of PKCδ and IBA1 in brain sections from vehicle- and niacin-treated mice. Statistical comparisons among multiple treatment groups were conducted using one-way ANOVA followed by Benjamini-Hochberg correction. Differences in Prkcd expression between spatial slides were assessed using the Wilcoxon rank-sum test ( p < 0.05). Data in (C and D), and I are presented as mean ± SEM. Scale bars on IF images: 50 μm.

Article Snippet: HMC3 – Human microglial cell line , ATCC , CRL-3304.

Techniques: In Vitro, Phagocytosis Assay, Labeling, Knockdown, Staining, Co-Culture Assay, Control, Comparison, Expressing

Journal: Bioactive Materials

Article Title: Bioengineering an improved three-dimensional vascularized co-culture model for studying Neuron–Microglia interactions

doi: 10.1016/j.bioactmat.2025.09.008

Figure Lengend Snippet: Differential impacts of distinct HMC3 microglial phenotypes on neuronal differentiation of hiNSCs within oriented SF scaffolds. (A) Schematic diagram for the construction of a neuron-microglia co-culture model on SF scaffold. (B) Immunofluorescence staining showing reduced neurite number and integrity in hiNSCs co-cultured with M1-polarized HMC3 cells, compared to M0, M2, and mono-culture groups. Scale bar, 100 μm. Quantification showing M1-polarized microglia impaired neuronal differentiation and network integrity in both scaffold surface (C) and central pore (D), while M2-polarized microglia preserved these features similar to mono-culture (n = 3). (E) RT-qPCR results showing reduced expression of neuronal markers ( SNAP-25 , GAP-43, SYP , and NFAT ) in hiNSCs co-cultured with HMC3 cells, most notably in the M1 group (n = 3). ∗P<0.05, ∗∗P<0.01, ∗∗∗P<0.001 .

Article Snippet: Human microglial cell line (HMC3, ATCC) was cultured in Minimum Essential Medium (MEM, Gibco) supplemented with 10 % fetal bovine serum (FBS, ExCell, Australia) and 1 % P/S (Solarbio), and maintained at 37 °C with 5 % CO 2 in a humidified incubator.

Techniques: Co-Culture Assay, Immunofluorescence, Staining, Cell Culture, Quantitative RT-PCR, Expressing