alpha mouse liver 12 aml12 hepatocyte cell line  (ATCC)

Journal: Journal of Extracellular Vesicles

Article Title: The Small Molecule H89 Facilitates Mesenchymal Stem Cell‐derived Extracellular Vesicle Release and Optimizes Therapeutic Efficacy in Liver Regeneration

doi: 10.1002/jev2.70285

Figure Lengend Snippet: H89 promotes extracellular vesicle (EV) production and release in hUCMSCs. (a) hUCMSCs were treated with 1 µM, 5 µM, or 10 µM H89 for 24 h, while cells in the control group received an equal volume of phosphate‐buffered saline (PBS). Immunofluorescence staining was used to visualize CD63 (red), and DAPI was used to stain nuclei (blue). The fluorescence intensity was quantified using ImageJ software ( n = 3). Scale bar: 100 µm. (b) Quantitative analysis of extracellular vesicle (EV) concentration by nanoflow cytometry (NanoFCM) in hUCMSCs pretreated with 10 µM H89 or mTORC1 inhibitors, as shown in Figure . The exact fold‐change for the H89 treatment group was 4.87‐fold compared to the control. (c) NanoFCM quantification of EV secretion by AML12 cells, HepRG cells, MSCs, and PTMSCs after 48 h of treatment with 10 µM H89. (d) Western blot analysis of CD63 and TSG101 expression in cellular lysates and EV fractions isolated from hUCMSCs treated with PBS (negative control) or 10 µM H89 for 48 h. (e, f) NanoFCM quantification of the expression of CD63, CD9, and CD81 in EVs, along with the particle size distribution and concentration. (g) EVs were visualized by transmission electron microscopy (TEM). EV density was quantified in 15 random fields ( n = 3). All the data are presented as the mean ± SD. Unpaired two‐tailed Student's t tests and One‐way ANOVA followed by Dunnett's multiple comparisons test were used to test for statistical significance. ** p < 0.01, *** p < 0.001.

Article Snippet: The murine hepatocyte cell line AML12 was purchased from Procell Life Science & Technology Co., Ltd. (Wuhan, China).

Techniques: Control, Saline, Immunofluorescence, Staining, Fluorescence, Software, Concentration Assay, Cytometry, Western Blot, Expressing, Isolation, Negative Control, Transmission Assay, Electron Microscopy, Two Tailed Test

Journal: Journal of Extracellular Vesicles

Article Title: The Small Molecule H89 Facilitates Mesenchymal Stem Cell‐derived Extracellular Vesicle Release and Optimizes Therapeutic Efficacy in Liver Regeneration

doi: 10.1002/jev2.70285

Figure Lengend Snippet: Modulation of the content of hUCMSC‐EVs by H89. (a) Proteomic analysis revealing the percentage of proteins that remained unchanged, were upregulated (>1.5‐fold), or were downregulated (<0.75‐fold) in H‐EVs compared with C‐EVs. (b) Selected differentially expressed proteins associated with “cell cycle regulation,” “energy metabolism,” and “oxidative stress,” along with their fold changes. (c) CCK‐8 assay demonstrating the effect of siRNA knockdown on AML12 cell proliferation. (d) Heatmap illustrating the changes in the expression of differentially expressed miRNAs between H‐EVs and C‐EVs. (e) Volcano plot highlighting the differentially expressed miRNAs. (f) The top 10 highly expressed miRNAs. (g) CCK‐8 assay to assess the effect of miRNA knockdown on AML12 cell proliferation. (h) GO and KEGG pathway analyses to identify the biological functions and pathways regulated by the target genes of miR‐29a. All the data are presented as the mean ± SD ( n = 3). Unpaired two‐tailed Student's t test was used to test for statistical significance. * p < 0.05, ** p < 0.01, ns: not significant.

Article Snippet: The murine hepatocyte cell line AML12 was purchased from Procell Life Science & Technology Co., Ltd. (Wuhan, China).

Techniques: CCK-8 Assay, Knockdown, Expressing, Two Tailed Test