Journal: Journal of Virology

Article Title: Pathogenic Old World Hantaviruses Infect Renal Glomerular and Tubular Cells and Induce Disassembling of Cell-to-Cell Contacts

doi: 10.1128/JVI.00568-11

Figure Lengend Snippet: Characterization of human renal cells and specificity of the anti-integrin antibodies. (A) HREpC, HRGEnC, and podocytes were analyzed for the presence or absence of the epithelial marker cytokeratin 18, the endothelial marker CD31, and the podocyte-specific protein synaptopodin, along with the specific isotype control antibodies. (B) Analysis of the specificity the of anti-integrin αVβ3 antibody LM609. Fixed renal cells were incubated with isotype control antibody, with anti-integrin αVβ3 LM609 or with anti-integrin αVβ3 antibody LM609 that was preincubated with recombinant human integrin αVβ3. (C) Specificity of the anti-integrin β3 antibody was confirmed by the absence of an integrin-specific band in K562 lysate lacking integrin expression (30).

Article Snippet: Human renal proximal epithelial cells (HREpC) were obtained from Promocell (Heidelberg, Germany) and maintained in renal epithelial cell growth medium 2 (Promocell).

Techniques: Marker, Incubation, Recombinant, Expressing

Journal: Journal of Virology

Article Title: Pathogenic Old World Hantaviruses Infect Renal Glomerular and Tubular Cells and Induce Disassembling of Cell-to-Cell Contacts

doi: 10.1128/JVI.00568-11

Figure Lengend Snippet: Epithelial barrier function is disturbed in hantavirus-infected cells. (A) HREpC were cultured on transwell filters and were infected with HTNV. Transepithelial resistance was measured to analyze the effect of infection on monolayer integrity. The initial TER of the uninfected and infected monolayers was set to 100%. Symbols: •, uninfected cells; ▾. HTNV-infected cells. Shown are data representative of three independent experiments (mean ± the SD). Student t test: *, P < 0.05; **, P < 0.01. (B) HREpC were infected with HTNV at an MOI of 0.01, and infection was monitored by the detection of N protein in cell lysate. Viability was assessed 144 h postinfection (hpi) by measuring the amount of ATP. Control cells remained uninfected. Viability of infected cells is presented as a percentage of uninfected cell control values. The data are representative of three independent experiments (mean ± the SD). Student t test, P=0.818.

Article Snippet: Human renal proximal epithelial cells (HREpC) were obtained from Promocell (Heidelberg, Germany) and maintained in renal epithelial cell growth medium 2 (Promocell).

Techniques: Infection, Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: Impact of High Glucose on Ocular Surface Glycocalyx Components: Implications for Diabetes-Associated Ocular Surface Damage

doi: 10.3390/ijms232214289

Figure Lengend Snippet: ( A ) Effect of a high glucose level (15 mM; 30 mM) on MUC 1, 4, and 16 gene expression in stratified human corneal epithelial cells. ( B ) Effect of a high glucose level (15 mM; 30 mM) on MUC 1, 4, and 16 gene expression in stratified human conjunctival epithelial cells. The data represent mean + S.E.M of cell culture experiments conducted in triplicate. * p < 0.05 compared to control cells exposed to normal (5 mM) glucose level.

Article Snippet: Human corneal epithelial cells were cultured in keratinocyte growth medium (PromoCell GmbH, Heidelberg, Germany) supplemented with bovine pituitary extract (0.004 mL/mL), human epidermal growth factor (0.125 ng/mL), human insulin (5 μg/mL), hydrocortisone (0.33 μg/mL), epinephrine (0.39 μg/mL), transferrin (10 μg/mL), and calcium chloride (0.15 mM).

Techniques: Expressing, Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: Impact of High Glucose on Ocular Surface Glycocalyx Components: Implications for Diabetes-Associated Ocular Surface Damage

doi: 10.3390/ijms232214289

Figure Lengend Snippet: ( A ) Effect of a high glucose level (15 mM; 30 mM) on MUC 1, 4, and 16 protein expression in stratified human corneal epithelial cells. ( B ) Effect of a high glucose level (15 mM; 30 mM) on MUC 1, 4, and 16 protein expression in stratified human conjunctival epithelial cells. The data represent mean + S.E.M of cell culture experiments conducted in triplicate.

Article Snippet: Human corneal epithelial cells were cultured in keratinocyte growth medium (PromoCell GmbH, Heidelberg, Germany) supplemented with bovine pituitary extract (0.004 mL/mL), human epidermal growth factor (0.125 ng/mL), human insulin (5 μg/mL), hydrocortisone (0.33 μg/mL), epinephrine (0.39 μg/mL), transferrin (10 μg/mL), and calcium chloride (0.15 mM).

Techniques: Expressing, Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: Impact of High Glucose on Ocular Surface Glycocalyx Components: Implications for Diabetes-Associated Ocular Surface Damage

doi: 10.3390/ijms232214289

Figure Lengend Snippet: ( A ) Representative brightfield images showing effect of a high glucose level (15 mM; 30 mM) on MUC 16 barrier function as measured by rose bengal exclusion assay in stratified human corneal epithelial cells. ( B ) Representative brightfield images showing effect of a high glucose level (15 mM; 30 mM) on MUC 16 barrier function as measured by rose bengal exclusion assay in stratified human conjunctival epithelial cells. Epithelial cells with potential loss of MUC 16 are stained pink with rose bengal. Graphs show the percent-stained area as quantified using Image J software of n = 6 images/group. The images were obtained from cell culture experiments conducted in triplicate. Scale bar = 100 μM.

Article Snippet: Human corneal epithelial cells were cultured in keratinocyte growth medium (PromoCell GmbH, Heidelberg, Germany) supplemented with bovine pituitary extract (0.004 mL/mL), human epidermal growth factor (0.125 ng/mL), human insulin (5 μg/mL), hydrocortisone (0.33 μg/mL), epinephrine (0.39 μg/mL), transferrin (10 μg/mL), and calcium chloride (0.15 mM).

Techniques: Exclusion Assay, Staining, Software, Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: Impact of High Glucose on Ocular Surface Glycocalyx Components: Implications for Diabetes-Associated Ocular Surface Damage

doi: 10.3390/ijms232214289

Figure Lengend Snippet: ( A ) Representative confocal microscopy images showing effect of a high glucose level (15 mM; 30 mM) on O-glycosylated Galβ1-3GalNAc oligosaccharide side chains of membrane-tethered mucins on stratified human corneal epithelial cells. ( B ) Representative confocal microscopy images showing effect of a high glucose level (15 mM; 30 mM) on O-glycosylated Galβ1-3GalNAc oligosaccharide side chains of membrane-tethered mucins on stratified human conjunctival epithelial cells. Nuclei are stained blue with DAPI, and O-glycosylated linked oligosaccharides are stained green with jacalin. Graphs show the percent-stained area as quantified using Image J software of n = 6 images/group. The images were obtained from cell culture experiments conducted in triplicate. Scale bar = 100 μM.

Article Snippet: Human corneal epithelial cells were cultured in keratinocyte growth medium (PromoCell GmbH, Heidelberg, Germany) supplemented with bovine pituitary extract (0.004 mL/mL), human epidermal growth factor (0.125 ng/mL), human insulin (5 μg/mL), hydrocortisone (0.33 μg/mL), epinephrine (0.39 μg/mL), transferrin (10 μg/mL), and calcium chloride (0.15 mM).

Techniques: Confocal Microscopy, Staining, Software, Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: Impact of High Glucose on Ocular Surface Glycocalyx Components: Implications for Diabetes-Associated Ocular Surface Damage

doi: 10.3390/ijms232214289

Figure Lengend Snippet: ( A ) Effect of a high glucose level (15 mM; 30 mM) on glycosyltransferase gene expression in stratified human corneal epithelial cells. ( B ) Effect of a high glucose level (15 mM; 30 mM) on glycosyltransferase gene expression in stratified human conjunctival epithelial cells. The data represent mean + S.E.M of cell culture experiments conducted in triplicate.

Article Snippet: Human corneal epithelial cells were cultured in keratinocyte growth medium (PromoCell GmbH, Heidelberg, Germany) supplemented with bovine pituitary extract (0.004 mL/mL), human epidermal growth factor (0.125 ng/mL), human insulin (5 μg/mL), hydrocortisone (0.33 μg/mL), epinephrine (0.39 μg/mL), transferrin (10 μg/mL), and calcium chloride (0.15 mM).

Techniques: Expressing, Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: Impact of High Glucose on Ocular Surface Glycocalyx Components: Implications for Diabetes-Associated Ocular Surface Damage

doi: 10.3390/ijms232214289

Figure Lengend Snippet: ( A ) Heat map showing changes in gene expression of proteins involved in proliferation and cell cycle in stratified human corneal epithelial cells exposed to high glucose concentration (15 mM; 30 mM). ( B ) Heat map showing changes in gene expression of proteins involved in proliferation and cell cycle in stratified human conjunctival epithelial cells exposed to high glucose concentration (15 mM; 30 mM). The proteins that show a fifty percent or higher decrease in expression are depicted in blue.

Article Snippet: Human corneal epithelial cells were cultured in keratinocyte growth medium (PromoCell GmbH, Heidelberg, Germany) supplemented with bovine pituitary extract (0.004 mL/mL), human epidermal growth factor (0.125 ng/mL), human insulin (5 μg/mL), hydrocortisone (0.33 μg/mL), epinephrine (0.39 μg/mL), transferrin (10 μg/mL), and calcium chloride (0.15 mM).

Techniques: Expressing, Concentration Assay