Journal: Cell Reports Medicine

Article Title: Ancestral library identifies conserved reprogrammable liver motif on AAV capsid

doi: 10.1016/j.xcrm.2022.100803

Figure Lengend Snippet: Library production and in vitro infectivity (A) A barcoded AAV vector library was produced either with (N = 5) or without (N = 3) addition of an exogenous assembly-activating protein (AAP). Individual dots in MA plots represent distinct barcodes, colors represent the amino acid identity at position 5. To the right of MA plots, eCDF plots with SEM depicted as horizontal error bars. (B) Results of an independent sites linear elastic net regularization approach show that addition of AAP modifies the impact of certain sites on production of vector. (C) Huh7 cells were transduced with the barcoded AAV library and DNA/RNA were isolated from those transduced cells (N = 5 per condition). Individual barcodes in MA plots are colored by identity at position 3 within the library. Extant barcoded vectors were also spiked into this transduction mixture. Fold change is plotted as bar graphs (error bars determined by bootstrapping, 1,000 replicates). (D) Regularization and linear modeling approach reveal potential similarities and differences among our positions of variation with respect to transduction and gene expression in vitro .

Article Snippet: Huh7 Cell Line , ATCC , PTA-4583.

Techniques: In Vitro, Infection, Plasmid Preparation, Produced, Transduction, Isolation, Gene Expression

Journal: Cell Reports Medicine

Article Title: Ancestral library identifies conserved reprogrammable liver motif on AAV capsid

doi: 10.1016/j.xcrm.2022.100803

Figure Lengend Snippet:

Article Snippet: Huh7 Cell Line , ATCC , PTA-4583.

Techniques: Virus, Plasmid Preparation, Recombinant, Imaging, RNA Sequencing, Software, Microscopy

Journal: Polymers

Article Title: Bioactive and Bioadhesive Catechol Conjugated Polymers for Tissue Regeneration

doi: 10.3390/polym10070768

Figure Lengend Snippet: Cell viability of human bone marrow mesenchymal stem cells (hBMSCs) treated with medium extracts of VHC films taken at different times. The diagrams include the mean and the standard deviation ( n = 8).

Article Snippet: Cellular toxicity and reactive oxygen species (ROS) assays were evaluated using human bone marrow mesenchymal stem cells (hBMSCs) (Innoprot, Derio, Vizcaya, Spain, P5), and anti-inflammatory activity was analyzed with murine RAW 264.7 macrophages (ECACC, Sigma-Aldrich, Saint Louis, MO, USA, P11). hBMSCs were cultured in DMEM enriched with 5% of fetal bovine serum (FBS), 5 mL of mesenchymal stem cell growth supplement (MSCGS), 50 μg/mL of Gentamicin (Sigma-Aldrich) and 2.5 μg/mL of Amphotericin B (Gibco, Waltham, MA, USA); and macrophages were cultured with DMEM enriched with sodium pyruvate (110 mg/L), 10% FBS, 100 μg/mL streptomycin, 100 units/mL penicillin and 200 mM l -glutamine.

Techniques: Standard Deviation

Journal: Polymers

Article Title: Bioactive and Bioadhesive Catechol Conjugated Polymers for Tissue Regeneration

doi: 10.3390/polym10070768

Figure Lengend Snippet: Intracellular reactive oxygen species (ROS) activity in hBMSCs measured from fluorescence emission at different times after treatment with VHC films extracts collected at 24 h. The diagrams include the mean, the standard deviation ( n = 4) and the analysis of variance (ANOVA) between the different groups and the positive control at each time (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Cellular toxicity and reactive oxygen species (ROS) assays were evaluated using human bone marrow mesenchymal stem cells (hBMSCs) (Innoprot, Derio, Vizcaya, Spain, P5), and anti-inflammatory activity was analyzed with murine RAW 264.7 macrophages (ECACC, Sigma-Aldrich, Saint Louis, MO, USA, P11). hBMSCs were cultured in DMEM enriched with 5% of fetal bovine serum (FBS), 5 mL of mesenchymal stem cell growth supplement (MSCGS), 50 μg/mL of Gentamicin (Sigma-Aldrich) and 2.5 μg/mL of Amphotericin B (Gibco, Waltham, MA, USA); and macrophages were cultured with DMEM enriched with sodium pyruvate (110 mg/L), 10% FBS, 100 μg/mL streptomycin, 100 units/mL penicillin and 200 mM l -glutamine.

Techniques: Activity Assay, Fluorescence, Standard Deviation, Positive Control

Journal: EMBO Molecular Medicine

Article Title: GP64-pseudotyped lentiviral vectors target liver endothelial cells and correct hemophilia A mice

doi: 10.1038/s44321-024-00072-8

Figure Lengend Snippet: ( A , B ) Mean and SEM with single values of the % ( A ) or mean fluorescent intensity (MFI, B ) of GFP-positive primary human hepatocytes ( n = 3 technical replicates) transduced with VSV.G-LV or GP64-LV at the indicated multiplicity of infections (MOI). Mann–Whitney test. ( C ) Mean and SEM with single values of the amount of hFIX measured in the culture supernatant of primary human hepatocytes ( n = 3 technical replicates) transduced with VSV.G-LV or GP64-LV at the indicated MOI, or left untreated. ( D ). Mean and SEM with single values of the vector copy numbers (VCN) of reversed transcribed LV genome of the primary human hepatocytes (shown in ( A – C ), n = 3 technical replicates) transduced with VSV.G-LV or GP64-LV encoding for GFP or hFIX at the indicated MOI, or left untreated. ( E , F ) Mean and SEM with single values of the % ( E ) or MFI ( F ) of GFP-positive primary human immortalized LSEC derived from a healthy donor ( n = 4 independent experiments) transduced with VSV.G-LV or GP64-LV at the indicated MOI. ( G ) Mean and SEM with single values of the amount of hFVIII measured in the culture supernatant of primary human immortalized LSEC derived from a healthy donor ( n = 4 independent experiments) transduced with VSV.G-LV or GP64-LV at the indicated MOI, or left untreated. ( H ) Mean and SEM with single values of the VCN of reversed transcribed LV genome of the primary human LSEC (shown in ( E – G )) transduced with VSV.G-LV or GP64-LV encoding for GFP or hFVIII at the indicated MOI, or left untreated. .

Article Snippet: Immortalized human LSEC were purchased from Innoprot (P10652-IM).

Techniques: Transduction, MANN-WHITNEY, Plasmid Preparation, Derivative Assay