human umbilical cord ecs (huvecs)  (Angiocrine)

Journal: eLife

Article Title: Human primed ILCPs support endothelial activation through NF-κB signaling

doi: 10.7554/eLife.58838

Figure Lengend Snippet: ( a ) Circulating human ILCs are identified as lineage negative CD127 + cells; within this population, we discriminate ILC1s as c-Kit - CRTH2 - , ILC2s as CRTH2 + c-Kit +/- , and ILCPs as c-Kit + CRTH2 - cells. HUVEC cells were co-cultured for 3 hr at 1:1 ratio in direct contact with either in vitro-expanded ( b ) or directly ex vivo-sorted ( c ) ILC1s, ILC2s, and ILCPs. Untreated ECs were employed as negative control (CTRL). ECs were harvested and analyzed for cell-surface adhesion molecule expression by flow cytometry. Graphs show representative histograms (panels b and c , top) and the summary (panels b and c , bottom) of the induction of the indicated adhesion molecules on the EC surface (n = 6). Ordinary one-way ANOVA–Tukey’s multiple comparison test (panel b ); Ordinary one-way ANOVA–Friedman test (panel c ). Figure 1—source data 1. Raw data of panels b and c.

Article Snippet: Primary human umbilical cord vein ECs (HUVECs – Lonza) and primary HDBECs (Promocell) were cultured in supplemented EC growth medium (EGM Ready To Use, Lonza) and used between passages 4 and 6.

Techniques: Cell Culture, In Vitro, Ex Vivo, Negative Control, Expressing, Flow Cytometry

Journal: eLife

Article Title: Human primed ILCPs support endothelial activation through NF-κB signaling

doi: 10.7554/eLife.58838

Figure Lengend Snippet: ( a ) The supernatant of the 3 hr co-culture experiments between ECs and ILCPs was analyzed for its cytokine contents (n = 4). The composition of the supernatant of ECs in EC growth medium was used as negative control (CTRL). ( b ) The expression of IL-6, IL-8, GM-CSF, TNF, and IFN-γ was analyzed by qPCR in ECs and ILCPs after being cultured for 3 hr at 1:1 ratio and FACS-sorted according to CD31 expression. Untreated ECs and ILCPs were employed as controls (CTRL). ( c ) HUVEC cells were co-cultured for 3 hr at 1:1 ratio in direct contact with in vitro-expanded ILCPs either in the absence (red dots) or presence (red circles) of a transwell (TW) insert (0.4 μm pore polycarbonate filter) or ( d ) in the presence of pre-conditioned media coming from previous EC–ILCP 3 hr co-cultures. ECs were harvested and analyzed for cell-surface adhesion molecule expression by flow cytometry (n = 6). The dotted lines indicate the level of average expression of adhesion molecules by unstimulated ECs. Statistical tests used: Unpaired t-test (panels a and d ); paired t-test (panel c ). Figure 2—source data 1. Raw data of panels a–d.

Article Snippet: Primary human umbilical cord vein ECs (HUVECs – Lonza) and primary HDBECs (Promocell) were cultured in supplemented EC growth medium (EGM Ready To Use, Lonza) and used between passages 4 and 6.

Techniques: Co-Culture Assay, Negative Control, Expressing, Cell Culture, In Vitro, Flow Cytometry