pe conjugated cd16  (Bio-Rad)

Journal: Journal of Tissue Engineering

Article Title: Blood mononuclear cells induce accelerated vascular remodeling under acute inflammation in vitro

doi: 10.1177/20417314251381716

Figure Lengend Snippet: Flow cytometry analysis of MCs collected before and after acute inflammation: (a) MCs’ analysis using CD14 and CD16 markers revealed the shift in surface marker intensity of a specific cell population at different stages of acute inflammation. (b) Cell-specific gating showed that primarily monocytes transited toward a CD14++/16+ state following acute inflammation. Interestingly, these cells marker intensity come to initial state after prolonged periods of acute inflammation. (c) The fraction of CD14++/16+ monocytes shows a profile similar to that of in vitro differentiated EC-like cells at different periods of acute inflammation. This implies that monocytes in CD14++/16+ state significantly contribute to the differentiation of EC-like cells in vitro . However, no significant intensity shift was observed in granulocytes or lymphocytes. In vitro treatment of PBMC with IL-6 also promotes monocyte transition toward a CD14++/16+ state and increases the EC-like cell fraction. (d) In vitro IL-6 treatment significantly enhances the population of CD34+, VEGFR2+, and CD163+ monocytes. Data were expressed as means ± standard deviations of one representative experiment out of three experiments carried out in triplicate (one-way ANOVA, Fraction ratio of monocytes: F 3,8 = 9.1, p = 0.0011; granulocytes: F 3,8 = 6.3, p = 0.0030; lymphocytes: F 3,8 = 1.2, p = 0.247, Tukey’s HSD post hoc test, * p < 0.05, ** p < 0.01, *** p < 0.001). Paired t-tests (n = 3) were used to evaluate differences among the groups in (d). Values of p < 0.05 are considered statistically significant, and are presented as follows: # p < 0.05.

Article Snippet: MCs and EC-like cells were stained with FITC-conjugated CD14 (MCA1218F), PE-conjugated anti-CD31 (MCA1746PET), and PE-conjugated CD16 (MCA1971PE) supplied by Bio-Rad Laboratories (Montreal, Quebec), and CD34 (bs-0646R-PE, Bioss Antibody Inc., Boston, MA, USA).

Techniques: Flow Cytometry, Marker, In Vitro

Journal: The Journal of Headache and Pain

Article Title: Expression of miR-155 in monocytes of people with migraine: association with phenotype, disease severity and inflammatory profile

doi: 10.1186/s10194-024-01842-y

Figure Lengend Snippet: Percentage of events recorded for the different monocytic sub-populations among the three study groups. Legend: Panel A : percentage of M1 monocytes (CD80 +) sub-populations. Panel B : percentage of M2 monocytes (CD163 +) sub-populations. CM-MO: chronic migraine with medication overuse, EM: episodic migraine, HCs: healthy controls, classical: “classical” monocytes (CD14 + /CD16– expression), non-classical: “non classical-intermediate” monocytes (CD14 + /CD16 + expression), M1: pro-inflammatory “M1” monocytes (CD80 + expression), M2 : anti-inflammatory “M2” monocytes (CD163 + expression). Box-plot: the range between the upper and lower border of the box indicates the interquartile range (IQR), spanning from the 25 th to the 75 th percentile. Within the box, the line indicates the median, and the cross denotes the mean. The upper and lower whiskers extend to the maximum and minimum values, excluding outliers. Symbols positioned above the upper whisker represent outliers, defined statistically as values beyond the 75 th percentile plus 1.5 times the IQR. Kruskal–Wallis Test was used for intergroup comparisons

Article Snippet: Subsequently, they were incubated for 30 min at 4 °C in the dark with all of the following monoclonal antibodies: Peridinin Chlorophyll Protein Complex (PerCP)-conjugated anti-human CD14 (BD Biosciences, 20µL per 1 × 10 6 cells), R-phyco-erythrin-cyanine7 (PE-Cy7)-conjugated anti-human CD16 (BD Biosciences, 5µL per 1 × 106 cells), and R-phycoerythrin (PE)-conjugated CD163 (BD Biosciences, 5µL per 1 × 106 cells) or R-phycoerythrin (PE)-conjugated CD80 (BD Biosciences, 5µL per 1 × 106 cells).

Techniques: Expressing, Whisker Assay