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: Materials Today Bio

Article Title: Vascular tissue reconstruction by monocyte subpopulations on small-diameter acellular grafts via integrin activation

doi: 10.1016/j.mtbio.2023.100847

Figure Lengend Snippet: The surface maker for the antibody used in this study.

Article Snippet: The cells were stained with PE-conjugated anti-CD31 antibody (MCA1746PET, Bio-Rad Laboratories, Montreal, Quebec), anti-CD34 antibody (bs-0646R-PE, Bioss Antibody Inc.), anti-CD105 antibody (bs-0579R-PE, Bioss Antibody Inc.), and anti-Flk-1 antibody (bs-0565R-PE, Bioss Antibody Inc), anti-CD163 antibody (bs-2527R-PE, Bioss Antibody Inc.), anti-CD14 antibody (MCA1218F, Bio-Rad Laboratories, Inc., Hercules, CA), and anti-CD16 antibody (MCA1971PE, Bio-Rad Laboratories, Inc.).

Techniques: Expressing, Marker

Journal: Materials Today Bio

Article Title: Vascular tissue reconstruction by monocyte subpopulations on small-diameter acellular grafts via integrin activation

doi: 10.1016/j.mtbio.2023.100847

Figure Lengend Snippet: Surface marker analysis of captured cells and porcine monocyte. The captured cells were isolated from the 3 h transplantation P-graft. a , Expression levels of CD31, CD34, CD105 and Flk-1 in CC-3H were indicated. b , Two-dimensional expression patterns of CD163/CD14 and CD16/CD14 in the captured cells were plotted. c , Two-dimensional expression patterns of CD163/CD14 and CD16/CD14 in the porcine monocyte were plotted. The MoN and MoP populations were expressed CD14 Low /CD16 Low /CD163 + and CD14 + /CD16 + /CD163 + , respectively.

Article Snippet: The cells were stained with PE-conjugated anti-CD31 antibody (MCA1746PET, Bio-Rad Laboratories, Montreal, Quebec), anti-CD34 antibody (bs-0646R-PE, Bioss Antibody Inc.), anti-CD105 antibody (bs-0579R-PE, Bioss Antibody Inc.), and anti-Flk-1 antibody (bs-0565R-PE, Bioss Antibody Inc), anti-CD163 antibody (bs-2527R-PE, Bioss Antibody Inc.), anti-CD14 antibody (MCA1218F, Bio-Rad Laboratories, Inc., Hercules, CA), and anti-CD16 antibody (MCA1971PE, Bio-Rad Laboratories, Inc.).

Techniques: Marker, Isolation, Transplantation Assay, Expressing

Journal: Materials Today Bio

Article Title: Vascular tissue reconstruction by monocyte subpopulations on small-diameter acellular grafts via integrin activation

doi: 10.1016/j.mtbio.2023.100847

Figure Lengend Snippet: Outgrowth capacity and surface marker analysis of captured cells a , Outgrowth capacity was evaluated on the cell culture plate. The cells were grown from the tissue after 3 days. b,c, Two-type cells were isolated from the single colony cultivation. The cells were classified into ( b) filopodia-shaped (CC-3D 1 ) and (c) spindle-shaped (CC-3D 2 ) cells based on the difference in morphology. These cells were expressed the surface maker of CD16/CD14. d , e , Surface marker expression of CD31, CD34, CD105, and Flk-1 on ( d ) CC-3D 1 and ( e ) CC-3D 2 were indicated. f , Di-ac-LDL uptake of the mixture of CC-3D 1 and CC-3D 2 was compared to the fibroblast and endothelial cells on CLSM observation.

Article Snippet: The cells were stained with PE-conjugated anti-CD31 antibody (MCA1746PET, Bio-Rad Laboratories, Montreal, Quebec), anti-CD34 antibody (bs-0646R-PE, Bioss Antibody Inc.), anti-CD105 antibody (bs-0579R-PE, Bioss Antibody Inc.), and anti-Flk-1 antibody (bs-0565R-PE, Bioss Antibody Inc), anti-CD163 antibody (bs-2527R-PE, Bioss Antibody Inc.), anti-CD14 antibody (MCA1218F, Bio-Rad Laboratories, Inc., Hercules, CA), and anti-CD16 antibody (MCA1971PE, Bio-Rad Laboratories, Inc.).

Techniques: Marker, Cell Culture, Isolation, Expressing

Journal: Materials Today Bio

Article Title: Vascular tissue reconstruction by monocyte subpopulations on small-diameter acellular grafts via integrin activation

doi: 10.1016/j.mtbio.2023.100847

Figure Lengend Snippet: In vitro differentiation assay on decellularized graft a , Schematic image of the culture plate with decellularized tissue and REDV-modified decellularized tissue. b , Two-dimensional plot showing the SSC and FSC signals of pre-seeding cells and cultured cells on the REDV-modified surface for 7 and 14 days. c, d , Surface marker analysis by FACS; the surface maker expressions of CD14 and CD16 are indicated. The blue line indicates the expression of the monocyte fraction. The black and red lines indicate the expression levels of the cells captured on the Un-graft and P-graft surfaces, respectively. d , Surface marker expression of CD31 and CD34 were indicated. In both cases, monocytes and cultured cells on decellularized and REDV-tissue surface for 7 and 14 days were evaluated. The gray-filled distribution indicates the unstained cells. The blue line indicates the antibody-stained monocyte. Black and red lines indicate the antibody-stained cells cultured on the unmodified and REDV-modified decellularized surfaces, respectively. e , Fraction rate of CD31 and CD34 in the cultured cells on unmodified and REDV-modified surfaces plotted against cultivation time. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The cells were stained with PE-conjugated anti-CD31 antibody (MCA1746PET, Bio-Rad Laboratories, Montreal, Quebec), anti-CD34 antibody (bs-0646R-PE, Bioss Antibody Inc.), anti-CD105 antibody (bs-0579R-PE, Bioss Antibody Inc.), and anti-Flk-1 antibody (bs-0565R-PE, Bioss Antibody Inc), anti-CD163 antibody (bs-2527R-PE, Bioss Antibody Inc.), anti-CD14 antibody (MCA1218F, Bio-Rad Laboratories, Inc., Hercules, CA), and anti-CD16 antibody (MCA1971PE, Bio-Rad Laboratories, Inc.).

Techniques: In Vitro, Differentiation Assay, Modification, Cell Culture, Marker, Expressing, Staining