primary cells  (InvivoGen)

Journal: The Cell Surface

Article Title: Compromising UPD-sugar nucleotide biosynthesis attenuates Candida albicans viability, virulence and drug sensitivity ☆

doi: 10.1016/j.tcsw.2026.100170

Figure Lengend Snippet: Host cell damage and virulence capacity of mutants in sugar nucleotide biosynthesis. (A) Mutants grown in +/− 25 μg/ml Dox screened for epithelial damage using A-431 cells by LDH assay. The mean LDH released at 24 h post co-incubation is shown for repressed mutants (grown in presence of Dox; blue bars) and No-Dox controls (red bars). Red and blue horizontal lines indicate the mean LDH activity for wild type control (No-Dox) and wild type grown in presence of Dox respectively. Welsh t-test used for statistical analysis; error bars represent standard error of mean; p**** < 0.0001. (B) Survival plots of G. mellonella larvae infected with C. albicans mutants in: (I) GDP-mannose, (II) UDP-glucose and (III) UDP- N -acetylglucosamine biosynthesis in presence (solid lines) and absence (dotted lines) of Dox. No killing or improved survival was observed for a number of repressed mutants. No killing was observed in control larvae injected with equivalent volume of PBS. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Human epithelial cells derived from a vulvar squamous cell carcinoma (A-431 cell line; ATCC No.: CRL-1555) were cultured and maintained in DMEM medium supplemented with 10% ( v /v) heat inactivated foetal calf serum, 5% penicillin and 5% streptomycin.

Techniques: Lactate Dehydrogenase Assay, Incubation, Activity Assay, Control, Infection, Injection

Journal: One Health

Article Title: Human 3D liver spheroids support productive infection of a novel tick-borne phenuivirus

doi: 10.1016/j.onehlt.2026.101321

Figure Lengend Snippet: Assembly and characterization of human 3D liver spheroids via DNA origami NAC-linkers. (A) Schematic of 3D liver spheroid self-assembly from primary human hepatocytes, liver sinusoidal endothelial cells, and Kupffer cells using NAC-linkers. (B) Atomic force microscopy image of NAC-linkers. Scale bars, 200 nm. (C) 1% agarose gel electrophoresis confirming cholesterol-modified NAC-linkers assembly (lanes: DNA marker, M13mp18 scaffold, and NAC-linkers). (D) Bright-field image of a mature spheroid. (E) Hematoxylin and eosin (H&E) staining of a spheroid section. (F) Immunofluorescence staining of cell type markers in human 3D liver spheroids: albumin (ALB, hepatocytes), CD31 (endothelial cells), and CD68 (Kupffer cells). Scale bars, 200 μm.

Article Snippet: Primary human hepatocytes, liver sinusoidal endothelial cells, and Kupffer cells (IxCell Biotechnology) were mixed at specific ratios and co-incubated with NAC-Linker A and B (Puheng Biomedicine, NAC001) to facilitate NAC structure formation on the cell surfaces.

Techniques: Microscopy, Agarose Gel Electrophoresis, Modification, Marker, Staining, Immunofluorescence

Journal: One Health

Article Title: Human 3D liver spheroids support productive infection of a novel tick-borne phenuivirus

doi: 10.1016/j.onehlt.2026.101321

Figure Lengend Snippet: Adaptation and pathogenesis of MKWV in human 3D liver spheroids. (A) Schematic of serial passaging of the HLJ1 strain in spheroids, yielding the adapted NAC-Org5 strain. (B, C) Viral RNA copies (B) and TCID₅₀ titers (C) across passages (P1-P5). (D) Bright-field image of spheroids infected with passage 5 (P5) virus, showing structural disruption. Scale bar, 100 μm. (E) Quantification of spheroid diameter post-infection. (F) Transmission electron micrographs of virions within cytoplasmic vesicles of infected spheroids. Scale bars: 1 μm (left), 200 nm (right). (G) Representative images and quantification of nuclei showing infection-induced cell death. Scale bar, 200 μm. (H) Western blot detecting cleaved caspase-3 in spheroids at 48 and 72 h post-infection (hpi). (I) Multiplex immunofluorescence showing NAC-Org5 tropism for CD31 + endothelial cells and CD68 + Kupffer cells, with weaker detection in ALB + hepatocytes. Scale bar, 200 μm. (J) Functional assessment of infected spheroids: ATP (viability), ALT/AST/LDH (damage), ALB/urea (synthetic function). (K) RT-qPCR analysis of pro-inflammatory cytokine mRNA expression, normalized to β-actin. Data are mean ± SD ( n = 5 biological replicates). * p < 0.05, ** p < 0.01.

Article Snippet: Primary human hepatocytes, liver sinusoidal endothelial cells, and Kupffer cells (IxCell Biotechnology) were mixed at specific ratios and co-incubated with NAC-Linker A and B (Puheng Biomedicine, NAC001) to facilitate NAC structure formation on the cell surfaces.

Techniques: Passaging, Infection, Virus, Disruption, Transmission Assay, Western Blot, Multiplex Assay, Immunofluorescence, Functional Assay, Quantitative RT-PCR, Expressing

Journal: One Health

Article Title: Human 3D liver spheroids support productive infection of a novel tick-borne phenuivirus

doi: 10.1016/j.onehlt.2026.101321

Figure Lengend Snippet: Pathogenicity of the NAC-Org5 strain in murine models. (A) Experimental schematic for intracranial (3-day-old) and intraperitoneal (3-week-old) inoculation of BALB/c mice. (B, C) Survival (B) and weight change (C) of suckling mice after NAC-Org5 infection. (D) Viral load in tissues and blood of suckling mice at 7 dpi. (E, F) Survival (E) and weight change (F) of 3-week-old mice. (G) Viral load in tissues and blood of 3-week-old mice at 7 dpi. Data are from 3 independent experiments. (H) Representative H& E -stained liver sections from 3-week-old mice at 7 and 15 dpi, showing inflammatory infiltrates and hepatocyte necrosis that resolves by 15 dpi. Scale bar, 100 μm. *** p < 0.001.

Article Snippet: Primary human hepatocytes, liver sinusoidal endothelial cells, and Kupffer cells (IxCell Biotechnology) were mixed at specific ratios and co-incubated with NAC-Linker A and B (Puheng Biomedicine, NAC001) to facilitate NAC structure formation on the cell surfaces.

Techniques: Infection, Staining

Journal: Materials Today Bio

Article Title: Injectable chitosan-based hydrogel via in situ gelation modulates the inflammatory microenvironment and facilitates minimally invasive repair of peripheral nerve injury

doi: 10.1016/j.mtbio.2026.102814

Figure Lengend Snippet: Pro-angiogenic and pro-migratory effects of hydrogels. Immunofluorescence staining shows blood vessel formation of HUVECs in LPS-macrophage condition medium (A). Scratch assays and Transwell migration assays of HUVECs (B) and L929 cells (C). Quantification of junctions (D), branches (E), wound closure percentage (F–G) and migrated cells (H–I). One-way ANOVA with Tukey's post hoc test and n = 3 for D-I; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, n.s. ANOVA, analysis of variance; CS, chitosan; IBU, ibuprofen; GP, genipin; MA, methacrylic anhydride; LPS, lipopolysaccharides; HUVEC, human umbilical vein endothelial cell; n.s., not significant.

Article Snippet: Mouse fibroblasts (L929, ATCC), human umbilical vein endothelial cells (HUVECs, ATCC), and mouse macrophages (RAW 264.7, ATCC) were provided by the Cell Bank of the Chinese Academy of Sciences.

Techniques: Immunofluorescence, Staining, Migration

Journal: Materials Today Bio

Article Title: Targeting FN1 to overcome gemcitabine resistance in gallbladder cancer: Mechanistic insights and an iRGD-modified PEG-PLGA nanoparticle delivery strategy

doi: 10.1016/j.mtbio.2026.102877

Figure Lengend Snippet: Effect of FN1 knockdown on the immunosuppressive microenvironment in GBC-SD/GEM cells. Note: (A) Schematic of immunosuppressive cells and cytokines detection in tumor tissues; (B) FCM analysis of Tregs infiltration levels in GBC-SD/GEM cell xenograft tissues in various mouse groups; (C) FCM analysis of M2 and M1 macrophage infiltration levels in GBC-SD/GEM cell xenograft tissues from different mouse groups; (D-E) RT-qPCR analysis of immunosuppressive factors expression in GBC-SD/GEM cell xenograft tissues from various mouse groups; (F) Schematic of in vitro co-culture of GEM-resistant GBC cells with THP-1 and CD4 + T cells; (G) FCM analysis of Tregs levels in CD4 + T cells after co-culture with GBC-SD/GEM cells; (H) FCM analysis of M2 and M1 macrophage levels in THP-1 cells post co-culture with GBC-SD/GEM cells; (I) RT-qPCR analysis of IL-10 or CSF-1 expression in CD4 + T or THP-1 cells after co-culture with GBC-SD/GEM cells. In B-E, ∗ indicates p < 0.05 compared to the sh-NC + GEM group, each group consisting of 6 mice; in G-I, ∗ indicates p < 0.05 compared to the sh-NC group, # indicates p < 0.05 compared to the oe-NC group, experiments repeated three times.

Article Snippet: Human CD4 + T cells were isolated and enriched from mature lymphocytes (hPBMCs, PCS-800-011, ATCC, USA) using a Human CD4 + T Cell Enrichment Kit (8804-6814-74, Thermo Fisher Scientific, USA). hPBMCs were incubated with the enrichment reagents for 30 min to capture CD4 + T cells.

Techniques: Knockdown, Quantitative RT-PCR, Expressing, In Vitro, Co-Culture Assay

Journal: Materials Today Bio

Article Title: Targeting FN1 to overcome gemcitabine resistance in gallbladder cancer: Mechanistic insights and an iRGD-modified PEG-PLGA nanoparticle delivery strategy

doi: 10.1016/j.mtbio.2026.102877

Figure Lengend Snippet: Impact of NPs delivering si-FN1 on drug resistance and immune cell infiltration in GBC-SD/GEM cells. Note: (A) Schematic of the experimental setup for studying the impact of NPs delivering si-FN1 on GBC GEM resistance; (B-C) RT-qPCR (B) and Western Blot (C) analysis of FN1 and PI3K pathway protein expression in GBC-SD/GEM cells treated with NPs (si-FN1) and iRGD-NPs (si-FN1); (D) CCK-8 assay assessing the viability changes in GBC-SD/GEM cells after treatment with NPs (si-FN1) and iRGD-NPs (si-FN1); (E) Clonogenic assay evaluating colony formation in various groups of GBC-SD/GEM and NOZ/GEM cells; (F) FCM analysis of apoptosis in GBC-SD/GEM cells across different groups; (G) FCM analysis of Tregs levels in CD4 + T cells after co-culture with GBC-SD/GEM cells; (H) FCM analysis of M2 and M1 macrophage levels in THP-1 cells after co-culture with GBC-SD/GEM cells; (I) RT-qPCR analysis of IL-10 or CSF-1 expression in CD4 + T or THP-1 cells co-cultured with GBC-SD/GEM cells. ∗ indicates p < 0.05 compared to the NPs (si-NC) group, # indicates p < 0.05 compared to the NPs (si-FN1) group, experiments repeated three times.

Article Snippet: Human CD4 + T cells were isolated and enriched from mature lymphocytes (hPBMCs, PCS-800-011, ATCC, USA) using a Human CD4 + T Cell Enrichment Kit (8804-6814-74, Thermo Fisher Scientific, USA). hPBMCs were incubated with the enrichment reagents for 30 min to capture CD4 + T cells.

Techniques: Quantitative RT-PCR, Western Blot, Expressing, CCK-8 Assay, Clonogenic Assay, Co-Culture Assay, Cell Culture