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Isolation of Viable Adipocytes and Stromal Vascular Fraction from Human Visceral Adipose Tissue Suitable for RNA Analysis and Macrophage Phenotyping
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JoVE 신문 의학
Isolation of Viable Adipocytes and Stromal Vascular Fraction from Human Visceral Adipose Tissue Suitable for RNA Analysis and Macrophage Phenotyping
DOI:

06:22 min

October 27, 2020

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Chapters

  • 00:00Introduction
  • 00:46Visceral Adipose Tissue Collection and Enzymatic Digestion
  • 02:10RNA Extraction from Mature Adipocytes and microRNA Quantification Assay
  • 03:16Characterization of Macrophage Subsets from Stromal Vascular Fraction
  • 04:42Results: Identification of Macrophages from Visceral Adipose Tissue
  • 06:02Conclusion

Summary

자동 번역

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.

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