Journal: bioRxiv

Article Title: “A novel Organ-Chip system emulates three-dimensional architecture of the human epithelia and allows fine control of mechanical forces acting on it.”

doi: 10.1101/2020.08.02.233338

Figure Lengend Snippet: Immuno-fluorescence images showing endothelial marker expression and distribution along the vascular channel. It is lined with a tight monolayer of lung microvascular endothelial cells expressing endothelial specific markers PECAM-1 (cyan), VE-cadherin (red) and VWF (green).Composite image of the spiraled section of the bottom channel evidences that full cell coverage occurs homogeneously even in the section with complex geometry (spiral area).

Article Snippet: Primary Epidermal Keratinocytes; Normal, Human, Neonatal Foreskin (HEKn) (ATCC ® : PCS-200-010TM), Primary Dermal Fibroblast Normal (HDFn) (ATCC ® : PCS-201-010TM) and Human Microvascular Endothelial Cell (ATCC ® : CRL-3243TM) were grown in T-75 culture flasks in an atmosphere of 5% CO 2 at 37°C according to the instructions provide by the manufacturers.

Techniques: Fluorescence, Marker, Expressing

Journal: Kidney International Reports

Article Title: Ex vivo C5b-9 Deposition Test to Monitor Complement Activity in Clinical and Subclinical Atypical Hemolytic Uremic Syndrome and in Transplantation-Associated Thrombotic Microangiopathy

doi: 10.1016/j.ekir.2024.04.022

Figure Lengend Snippet: C5b-9 deposition area on resting and ADP-activated endothelial cells incubated with serum of patients with aHUS. C5b-9 deposition upon cultured endothelial cells induced by serum of patients with aHUS HMEC-1 cells were treated with ADP 10 μM or with dilution vehicle (mqH 2 O) for 10 minutes, and afterward with serum of the included patients for 4 hours. C5b-9 was detected by immunocytochemistry using a specific antibody. (a) C5b-9 staining area upon HMEC-1 cells induced by serum of the patients with aHUS included, calculated from 15 pictures in each case. The dotted rectangles represent the normal range of C5b-9 deposition. Data were analyzed using 2-way analysis of variance followed by Dunnet’s multiple comparison test (∗, P < 0.05 vs. respective control). (b) Representative images of C5b-9 deposits on HMEC-1 cells (in red) induced by serum of healthy controls (Ctl) and of the sample 1 and sample 2 of patient 4 (P4 S1 and P4 S2, respectively) in resting and in ADP-activated conditions. Nuclei were stained with Hoechst 33342 and are shown in blue. (c) Representative images of C5b-9 deposits on HMEC-1 cells (in red) induced by serum of healthy controls (Ctl) and of the sample 1 and sample 2 of patient 8 (P8 S1 and P8 S2, respectively) in resting and in ADP-activated conditions. Nuclei were stained with Hoechst 33342 and are shown in blue. The mean ± SD values of C5b-9 staining area in each case can be found in Supplementary Table S4. ADP, adenosine diphosphate; aHUS, atypical hemolytic uremic syndrome; HMEC-1, human dermal microvascular endothelial cells.

Article Snippet: We studied the C5b-9 deposition on human dermal microvascular endothelial cells (HMEC-1; ATCC CRL-3234) by immunofluorescence using a specific antibody against complement C5b-9 (204903, Calbiochem) as described by Noris et al.

Techniques: Incubation, Cell Culture, Immunocytochemistry, Staining, Comparison, Control

Journal: Kidney International Reports

Article Title: Ex vivo C5b-9 Deposition Test to Monitor Complement Activity in Clinical and Subclinical Atypical Hemolytic Uremic Syndrome and in Transplantation-Associated Thrombotic Microangiopathy

doi: 10.1016/j.ekir.2024.04.022

Figure Lengend Snippet: C5b-9 deposition area on resting and ADP-activated endothelial cells incubated with serum of patients with TMA secondary to kidney transplantation (KT) or lung transplantation (LT). HMEC-1 cells were treated with ADP 10μM or with dilution vehicle (mqH 2 O) for 10 minutes and, afterward with serum of the included patients for 4 hours. C5b-9 was detected by immunocytochemistry using a specific antibody. Representative images of C5b-9 deposits on HMEC-1 cells (in red) induced by serum of patients with kidney transplant–associated TMA (KT1 and KT2) and patients with lung transplant–associated TMA in resting and in ADP-activated conditions and the respective C5b-9 staining area calculated from 15 pictures in each case. The dotted rectangles represent the normal range of C5b-9 deposition. Data were analyzed using 2-way analysis of variance followed by Dunnet’s multiple comparison test (∗, P < 0.05 vs. respective control). The mean ± SD values of C5b-9 staining area in each case can be found in Supplementary Table S4. ADP, adenosine diphosphate; HMEC-1, human dermal microvascular endothelial cells; TMA, thrombotic microangiopathy.

Article Snippet: We studied the C5b-9 deposition on human dermal microvascular endothelial cells (HMEC-1; ATCC CRL-3234) by immunofluorescence using a specific antibody against complement C5b-9 (204903, Calbiochem) as described by Noris et al.

Techniques: Incubation, Transplantation Assay, Immunocytochemistry, Staining, Comparison, Control

Journal: Kidney International Reports

Article Title: Ex vivo C5b-9 Deposition Test to Monitor Complement Activity in Clinical and Subclinical Atypical Hemolytic Uremic Syndrome and in Transplantation-Associated Thrombotic Microangiopathy

doi: 10.1016/j.ekir.2024.04.022

Figure Lengend Snippet: C5b-9 deposition area on resting and ADP-activated endothelial cells incubated with serum of patients with TMA hematopoietic stem cell transplantation (HSCT). HMEC-1 cells were treated with ADP 10μM or with dilution vehicle (mqH 2 O) for 10 minutes and, afterward with serum of the included patients for 4 hours. C5b-9 was detected by immunocytochemistry using a specific antibody. Representative images of C5b-9 deposits on HMEC-1 cells (in red) induced by serum of patients with HSCT-associated TMA (HSCT 1 and 2) in resting and in ADP-activated conditions and the respective C5b-9 staining area calculated from 15 pictures in each case. The dotted rectangles represent the normal range of C5b-9 deposition. Data were analyzed using 2-way analysis of variance followed by Dunnet’s multiple comparison test (∗, P < 0.05 vs. respective control). The mean ± SD values of C5b-9 staining area in each case can be found in Supplementary Table S4. ADP, adenosine diphosphate; HMEC-1, human dermal microvascular endothelial cells; TMA, thrombotic microangiopathy.

Article Snippet: We studied the C5b-9 deposition on human dermal microvascular endothelial cells (HMEC-1; ATCC CRL-3234) by immunofluorescence using a specific antibody against complement C5b-9 (204903, Calbiochem) as described by Noris et al.

Techniques: Incubation, Transplantation Assay, Immunocytochemistry, Staining, Comparison, Control

Journal: Kidney International Reports

Article Title: Ex vivo C5b-9 Deposition Test to Monitor Complement Activity in Clinical and Subclinical Atypical Hemolytic Uremic Syndrome and in Transplantation-Associated Thrombotic Microangiopathy

doi: 10.1016/j.ekir.2024.04.022

Figure Lengend Snippet: An asymptomatic 7-year-old boy at risk of aHUS presented increased C5b-9 deposition on cultured endothelial cells during a persistent otitis episode. (a) Scheme of CFHR3-CFHR1 deletion inheritance (in blue). (b) Representative images of C5b-9 deposits on HMEC-1 cells (in red) induced by serum of healthy controls (Ctl) and of the patient in resting and in ADP-activated conditions. Nuclei were stained with Hoechst 33342 and are shown in blue. (c) C5b-9 staining area upon HMEC-1 cells induced by serum of the patient calculated from 15 pictures in each case. The dotted rectangles represent the normal range of C5b-9 deposition. Data were analyzed using 2-way analysis of variance followed by Dunnet’s multiple comparison test (∗, P < 0.05 vs. respective control) (d) Evolution of serum C3 and hematuria during the otitis episode of the patient. Point 0 represents the time when the serum sample for C5b-9 deposition test was collected. The evolution of the rest of clinical and complement data can be found in Supplementary Table S1. ADP, adenosine diphosphate; aHUS, atypical hemolytic uremic syndrome; HMEC-1, human dermal microvascular endothelial cells.

Article Snippet: We studied the C5b-9 deposition on human dermal microvascular endothelial cells (HMEC-1; ATCC CRL-3234) by immunofluorescence using a specific antibody against complement C5b-9 (204903, Calbiochem) as described by Noris et al.

Techniques: Cell Culture, Staining, Comparison, Control