Multicolor Flow Cytometry Analysis of Adipose Cells

Multicolor flow cytometry measurements were performed and analyzed as described previously [23 (link),25 (link)]. Briefly, cells isolated from the SVF, which were obtained following collagenase digestion and lysis of red blood cells, were stained for flow cytometry using two antibody cocktails. Cocktail 1 included CD45-PE-Cy7 (BD 557748), CD3-fitc (BD 561807), CD19-fitc (BD 555412), CD56-fitc (BD 562794), CD66b-fitc (BD 555724), CD11b-BV421 (Biologend 301324), and CD11c-APC-Cy7 (Biolegend 337218). Cocktail 2 included CD45-PE-Cy7 (BD 557748), CD3-V500 (BD 561416), CD4-PerCP (Biolegend 300528), CD8 APC-H7 (BD 641400), CD19-BV421 (Biolegend 302234), and CD56-APC (Biolegend 318310). Samples were measured with a FACS-Canto II (BD Biosciences, Gurugram, India). Results were analyzed with FACSdiva (BD Biosciences) and FlowJo software version 10. Since weight of the adipose tissues was unavailable, data are expressed as % of cells relative to total cells (based on forward and side scatter plot).

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De Hert E., Verboven K., Wouters K., Jocken J.W, & De Meester I. (2022). Prolyl Carboxypeptidase Activity Is Present in Human Adipose Tissue and Is Elevated in Serum of Obese Men with Type 2 Diabetes. International Journal of Molecular Sciences, 23(21), 13529.

Publication 2022

CD45-PE-Cy7 CD3-fitc CD19-fitc CD56-fitc CD66b-fitc CD11c-APC-Cy7 CD3-V500 CD4-PerCP CD8 APC-H7 CD19-BV421 CD56-APC FACS-Canto II FACSdiva

Adipose tissues Antibody cocktails Cd11b Cd66b Cells Collagenase Digestion Fitc Flow cytometry Red blood cells

Corresponding Organization : University of Antwerp

Other organizations : Hasselt University, Maastricht University Medical Centre

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Percentages of different cell types (CD45+, CD3+, CD19+, CD56+, CD66b+, CD11b+, CD11c+, CD4+, CD8+, CD19+, CD56+) relative to total cells

control variables

None explicitly mentioned

controls

Positive control: None mentioned
Negative control: None mentioned

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