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

Journal: iScience

Article Title: A human cornea-on-a-chip for the study of epithelial wound healing by extracellular vesicles

doi: 10.1016/j.isci.2022.104200

Figure Lengend Snippet: Schematic diagram of human cornea and design of the cornea-chip (A) The anatomy of the human cornea. (B) Cross-sectional schematic diagram of the corneal chip. HCEpi and HCEnd cells are cultured on the opposite sides of a porous PC membrane coated with ECM. The membrane is sandwiched between two PDMS layers incorporated with microfluidic channels. (C) Photographs of the human cornea-chip; upper and lower microfluidic structures are indicated by using solutions with purple and green color dye. (D) Three-dimensional illustration on the device, showing the details on the structures on different layers. (E) The cornea-chip with two individual microfluidic channels and a circular hole in the middle mimics the human corneal structure. Structures for in situ TEER measurements are also fabricated on the chip. (F) Experimental setup for flow control and cell culture. The device is located in an incubator with two microchannels connected to respective syringe pumps.

Article Snippet: Immortalized HCEnd cells , Applied Biological Materials Inc. , T5077.

Techniques: Cell Culture, Membrane, In Situ, Control

Journal: iScience

Article Title: A human cornea-on-a-chip for the study of epithelial wound healing by extracellular vesicles

doi: 10.1016/j.isci.2022.104200

Figure Lengend Snippet: Construction of biomimetic coculture platform of the human corneal cells in the microfluidic chip (A) Microscopy images on the cell viability of the HCEpi cells and the HCEnd cells cocultivated in the cornea-chip. Cells are treated by Calcein-AM (live cells, green color) and PI (dead cells, red color) before observation. (B) TEER values obtained from the cornea-chip at 3, 7, 10, and 14 days. (C) Measurement of the corneal permeability coefficient (Papp) on 5 kDa FITC-Dextran. (B) (C) All data are mean ± SD from three independent experiments.

Article Snippet: Immortalized HCEnd cells , Applied Biological Materials Inc. , T5077.

Techniques: Microscopy, Permeability

Journal: iScience

Article Title: A human cornea-on-a-chip for the study of epithelial wound healing by extracellular vesicles

doi: 10.1016/j.isci.2022.104200

Figure Lengend Snippet: Construction of physiological phenotypes of the human cornea (A and C) Immunofluorescent characterization of human corneal cells cultured in different conditions. "Epi only" or "End only" represents a single type of cells cultured in the chip, respectively. "Chip" means the coculture of the HCEpi cells and HCEnd cells. The green color is the immunofluorescence staining of ZO-1 protein, and the blue color shows the cell nucleus by DAPI staining. (B) Mean fluorescent intensity (MFI) of ZO-1 protein expression obtained from Figure 4 A. (D) MFI of ZO-1 protein expression obtained from Figure 4 C a.u., arbitrary unit. (B and D) Data are mean ± SD of relative fluorescence intensity from three independent experiments. P values by two-way ANOVA and Bonferroni’s multiple comparisons test. “ns” no significant difference, "∗∗" p < 0.01, "∗∗∗∗" p < 0.001.

Article Snippet: Immortalized HCEnd cells , Applied Biological Materials Inc. , T5077.

Techniques: Cell Culture, Immunofluorescence, Staining, Expressing, Fluorescence

Journal: iScience

Article Title: A human cornea-on-a-chip for the study of epithelial wound healing by extracellular vesicles

doi: 10.1016/j.isci.2022.104200

Figure Lengend Snippet:

Article Snippet: Immortalized HCEnd cells , Applied Biological Materials Inc. , T5077.

Techniques: Recombinant, Staining, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Software

Journal: Scientific Reports

Article Title: Single and fractionated ionizing radiation induce alterations in endothelial connexin expression and channel function

doi: 10.1038/s41598-019-39317-9

Figure Lengend Snippet: The effect of single and fractionated irradiation on Cx37, Cx40 and Cx43 gene expression. Gene expression of Cx37, Cx40 and Cx43 at 6 h, 24 h, 48 h, 72 h, 7 d or 14 d after a single X-ray exposure (0.1, 0.5 and 5 Gy) in TICAE (left side) and TIME cells (right side) (a,b,c). Gene expression of Cx37, Cx40 and Cx43 at 24 h (d) and 7 d (e) after single or fractionated irradiation in TICAE (left side) and TIME cells (right side). Fractionated irradiation involved three consecutive X-rays doses (0.033 and 1.67 Gy/fraction/day), leading to cumulative doses of 0.1 and 5 Gy. Data were analyzed with a nonparametric Mann-Whitney T-test. Values represent average ± SEM of 5 biological replicates, except for 6 h p.i. where 4 biological replicates were used. ( a – c ) *Indicates for a given time point the statistical difference of gene expression after a dose of single irradiation compared to the respective normalized 0 Gy controls at the same time point. ( d – e ) • Indicates for a given time point the statistical difference of gene expression after a dose of fractionated irradiation compared to the respective normalized 0 Gy controls at the same time point. ( d – e ) *Indicates the statistical difference between fold changes of gene expression after a given radiation dose and a given time of single and fractionated irradiation compared to the respective normalized 0 Gy controls at the same time point. */• p < 0.05; **/•• p < 0.01; ***/••• p < 0.0001. Cx, connexin; TICAE, Telomerase Immortalized human Coronary Artery Endothelial cells; TIME, Telomerase Immortalized human Microvascular Endothelial cells; p.i, post irradiation; h, hours; d, days; SEM, standard error of mean.

Article Snippet: We used two human endothelial cell lines: hTERT telomerase immortalized human coronary artery endothelial cells (TICAE) from the European Collection of Authenticated Cell Cultures (ECACC; HCAECs Cat. No: 300-05), and telomerase immortalized human dermal microvascular endothelial cells (TIME) from the American Type Cell Culture (ATTC).

Techniques: Irradiation, Gene Expression, MANN-WHITNEY