Visceral Adipose Tissue Collection and Enzymatic Digestion
2:10
RNA Extraction from Mature Adipocytes and microRNA Quantification Assay
3:16
Characterization of Macrophage Subsets from Stromal Vascular Fraction
4:42
Results: Identification of Macrophages from Visceral Adipose Tissue
6:02
Conclusion
Transcript
This protocol allows for isolation and study of viable adipocytes and stromal vascular fraction SVF cells from human visceral fat. Enzymatic digestion using collagenase ensures high yields of total RNA from mature adipocytes, including microRNAs s
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This protocol provides an efficient collagenase digestion method for isolation of viable adipocytes and stromal vascular fraction-SVF cells from human visceral fat in a single process, including methodology to obtain high-quality RNA from adipocytes and phenotypification of SVF-macrophages through staining of multiple membrane-bound markers for analysis by flow cytometry.