Journal: Toxicology Reports

Article Title: Genotoxicity risk assessment of a 7-hydroxymitragynine-enriched Kratom preparation: An integrated in silico and in vitro approach

doi: 10.1016/j.toxrep.2026.102206

Figure Lengend Snippet: Micronucleus test of Kratom leaf extract after 4 h exposure with S9 in TK6 cells. Results are the mean ± SD of 3 independent experiments. Statistical testing with one-way ANOVA and Tukey’s post-hoc test (* p < 0.05).

Article Snippet: The human B lymphoblastoid cell line (TK6) (CRL-8015; batch No. 70045146), purchased from ATCC, was cultured in RPMI 1640 medium supplemented with 10 % fetal bovine serum (FBS) and 1 % penicillin/streptomycin.

Techniques:

Journal: Toxicology Reports

Article Title: Genotoxicity risk assessment of a 7-hydroxymitragynine-enriched Kratom preparation: An integrated in silico and in vitro approach

doi: 10.1016/j.toxrep.2026.102206

Figure Lengend Snippet: Micronucleus test of Kratom leaf extract after 4 h exposure without S9 in TK6 cells. Results are the mean ± SD of 3 independent experiments. Statistical testing with one-way ANOVA and Tukey’s post-hoc test (* p < 0.05).

Article Snippet: The human B lymphoblastoid cell line (TK6) (CRL-8015; batch No. 70045146), purchased from ATCC, was cultured in RPMI 1640 medium supplemented with 10 % fetal bovine serum (FBS) and 1 % penicillin/streptomycin.

Techniques:

Journal: Toxicology Reports

Article Title: Genotoxicity risk assessment of a 7-hydroxymitragynine-enriched Kratom preparation: An integrated in silico and in vitro approach

doi: 10.1016/j.toxrep.2026.102206

Figure Lengend Snippet: Micronucleus test of Kratom leaf extract after 24 h exposure without S9 in TK6 cells. Results are the mean ± SD of 3 independent experiments. Statistical testing with one-way ANOVA and Tukey’s post-hoc test (* p < 0.05).

Article Snippet: The human B lymphoblastoid cell line (TK6) (CRL-8015; batch No. 70045146), purchased from ATCC, was cultured in RPMI 1640 medium supplemented with 10 % fetal bovine serum (FBS) and 1 % penicillin/streptomycin.

Techniques:

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: Bioactive Materials

Article Title: Multimodal profiling of CAR T cells against glioblastoma using a microengineered 3D tumor-on-a-chip model

doi: 10.1016/j.bioactmat.2026.01.003

Figure Lengend Snippet: Off-target cytotoxicity evaluation of CAR T cells using the 3D GOC system. A) Schematic representation of the differing cytolytic mechanisms of UTD, TV-13, and IL-13 CAR T cells against IL13Rα1 + HT-1080 tumor cells. Created with BioRender.com . B) Flow cytometric analysis confirming IL13Rα1 and mCherry (reporter gene) expression on IL13Rα1 + HT-1080 tumor cells. Antigen expression (IL13Rα1 or mCherry) on viable tumor cells shown in histograms: blue for IL13Rα1 + HT-1080 tumor cells and red for control tumor cells. The values within each histogram indicate the percentage of positive cells, with the mean fluorescence intensity (MFI) shown in parentheses. C) Microfluidic evaluation of off-target toxicities of T cells. (i) Representative tile images of tumor-stroma interface stained for actin cytoskeleton (green), showing differences in migration of IL13R1 + HT-1080 tumor cells (red) within the 3D GOC model across varying densities of UTD, TV-13 CAR, and IL-13 CAR T cells. (ii) Quantification of the migration distance of the IL13Rα1 + HT-1080 tumor cells in response to varying T cell concentrations. Data are represented as mean ± SD measured from three biological replicates ( n = 3) , T cell donors: DN18, DN28, and DN31, ∗ p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗ ∗p < 0.0001. Two-way ANOVA with Tukey's multiple comparisons test was utilized for statistical analysis. (iii) Bar graph showing the difference in nuclei per field of view (FOV) across different T cell densities, used as a measure of chain migration by IL13Rα1 + HT-1080 tumor cells. Data are represented as mean ± SD measured from three biological replicates ( n = 3) , T cell donors: DN18, DN28, and DN31, ∗ p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. Two-way ANOVA with Tukey's multiple comparisons test was utilized for statistical analysis, and (iv) Bar graph representing the percentage of T cells positive for intracellular cytokines in the presence of IL13Rα1 + HT-1080 tumor cells. Data are represented as mean ± SD measured from three biological replicates ( n = 3) , ∗ p < 0.05, ∗∗p < 0.01, ∗∗∗ ∗p < 0.0001. Two-way ANOVA with Tukey's multiple comparisons test was utilized for statistical analysis.

Article Snippet: HT-1080 Culture : Human fibrosarcoma cells (CCL-121, ATCC or HT-1080) were used to generate an off-target cell line (IL13Rα1 + HT-1080) expressing IL13Rα1-T2A-mCherry gene, which was single-sorted for the experiments described here.

Techniques: Gene Expression, Expressing, Control, Fluorescence, Staining, Migration

Journal: Neural Regeneration Research

Article Title: R-28 cell-derived extracellular vesicles protect retinal ganglion cells in glaucoma

doi: 10.4103/NRR.NRR-D-24-00709

Figure Lengend Snippet: In vitro testing of R-28 cell-derived EVs on RGC survival and regeneration. Representative images of untreated control wells (A), R-28 cell-derived EVs treated wells (B), and CNTF-treated wells (C) are shown with the graphs showing the total surviving RGC number (D), the number of RGC with neurites (E), and the longest neurite length (F) in primary retinal cell culture after 3 days. Data are expressed as the mean ± SEM. Images were stained with a nuclear (DAPI, blue) and RGC marker (β-III tubulin, green). Scale bars: 50 µm. All experiments were performed in three independent biological replicates. CNTF: Ciliary neurotrophic factor; DAPI: 4′,6-diamidino-2-phenylindole; EV: extracellular vesicles; RGC: retinal ganglion cells.

Article Snippet: To test the therapeutic effect of R-28-derived EVs on human ESC-derived RGC (H7/H9 immortalized cell line; WiCell, Madison, WI, USA, #WA07, RRID: CVCL_S800) were differentiated from CRISPR-modified ESC generously donated from Prof Donald Zacks laboratory (Johns Hopkins University, Baltimore, MD, USA) and licensed for use from WiCell (Material Transfer Agreement issue-164634007).

Techniques: In Vitro, Derivative Assay, Control, Cell Culture, Staining, Marker

Journal: Neural Regeneration Research

Article Title: R-28 cell-derived extracellular vesicles protect retinal ganglion cells in glaucoma

doi: 10.4103/NRR.NRR-D-24-00709

Figure Lengend Snippet: R-28 cell-derived EVs promote human ESC-derived RGC survival in vitro . Images show untreated controls, R-28 cell-derived EV treated, and CNTF-treated hESC-derived RGCs (green, βIII-tubulin) after injury induced by the microtubule poison, colchicine. Scale bar: 50 µm. Data are presented as mean ± SEM. All experiments were performed in three independent biological replicates. CNTF: Ciliary neurotrophic factor; ESC: embryonic stem cells; EV: extracellular vesicles; RGC: retinal ganglion cells.

Article Snippet: To test the therapeutic effect of R-28-derived EVs on human ESC-derived RGC (H7/H9 immortalized cell line; WiCell, Madison, WI, USA, #WA07, RRID: CVCL_S800) were differentiated from CRISPR-modified ESC generously donated from Prof Donald Zacks laboratory (Johns Hopkins University, Baltimore, MD, USA) and licensed for use from WiCell (Material Transfer Agreement issue-164634007).

Techniques: Derivative Assay, In Vitro

Journal: Neural Regeneration Research

Article Title: R-28 cell-derived extracellular vesicles protect retinal ganglion cells in glaucoma

doi: 10.4103/NRR.NRR-D-24-00709

Figure Lengend Snippet: R-28 cell-derived EVs show protective trend for RGCs in a chronic glaucoma model. (A) Experimental design of the in vivo study. R-28 cell-derived EVs were intravitreally injected weekly beginning 1 week after microbead injection, and animals’ IOPs were measured twice a week. Four weeks after weekly EV injection, animals were sacrificed and histologically analyzed. After injection, microbeads localized (arrow) around the iridocorneal angle (B). IOP (mmHg) of healthy animals (blue) and animals receiving intracameral injection of microbeads with (green) or without (brown) intravitreal EV treatments is shown (C). (D, E) Representative images (D) and quantification (E) of Brn3a + (green) RGCs from the three groups on week 5. Scale bars: 50 µm. Data are presented as mean ± SEM. n = 3–5. EV: Extracellular vesicles; IOP: intraocular pressure; RGC: retinal ganglion cells; PBS: phosphate buffered saline.

Article Snippet: To test the therapeutic effect of R-28-derived EVs on human ESC-derived RGC (H7/H9 immortalized cell line; WiCell, Madison, WI, USA, #WA07, RRID: CVCL_S800) were differentiated from CRISPR-modified ESC generously donated from Prof Donald Zacks laboratory (Johns Hopkins University, Baltimore, MD, USA) and licensed for use from WiCell (Material Transfer Agreement issue-164634007).

Techniques: Derivative Assay, In Vivo, Injection, Saline

Journal: Neural Regeneration Research

Article Title: R-28 cell-derived extracellular vesicles protect retinal ganglion cells in glaucoma

doi: 10.4103/NRR.NRR-D-24-00709

Figure Lengend Snippet: Differentially expressed miRNA shown as abundance and fold change heat map profiles. Heatmaps show the upregulated and downregulated normalized counts of miRNA from injured RGCs treated with R-28 cell-derived EVs compared to injured untreated (A, B), injured RGCs treated with R-28 derived EVs compared to uninjured treated (D, E), and uninjured RGCs treated with R-28 cell-derived EVs compared to injured untreated (G, H), both statistically significant ( P < 0.05; A, D, G) and those trending towards significance ( P < 0.1; B, E, H) with abundance profiles shown in associated bar charts (C, F, I, respectively). * P < 0.05, ** P < 0.01. Data are presented as mean ± SEM. n = 3. EV: Extracellular vesicles; RGC: retinal ganglion cells.

Article Snippet: To test the therapeutic effect of R-28-derived EVs on human ESC-derived RGC (H7/H9 immortalized cell line; WiCell, Madison, WI, USA, #WA07, RRID: CVCL_S800) were differentiated from CRISPR-modified ESC generously donated from Prof Donald Zacks laboratory (Johns Hopkins University, Baltimore, MD, USA) and licensed for use from WiCell (Material Transfer Agreement issue-164634007).

Techniques: Derivative Assay