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从人内脏脂肪组织中分离出活的脂肪细胞和基质血管组分，适用于 RNA 分析和巨噬细胞表型分析
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JoVE Journal Medicine
Isolation of Viable Adipocytes and Stromal Vascular Fraction from Human Visceral Adipose Tissue Suitable for RNA Analysis and Macrophage Phenotyping
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从人内脏脂肪组织中分离出活的脂肪细胞和基质血管组分，适用于 RNA 分析和巨噬细胞表型分析

DOI:
10.3791/61884-v

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06:22 min

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October 27, 2020

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, , , , , , , ,
1Research Division,Instituto Nacional de Perinatologia, 2Department of Immunobiochemistry,Instituto Nacional de Perinatologia, 3Department of Obstetrics,Instituto Nacional de Perinatologia, 4Department of Academic Programs and Continuing Education,Instituto Nacional de Perinatologia, 5Department of Infectology and Immunology,Instituto Nacional de Perinatologia, 6Clinical Research Branch,Instituto Nacional de Perinatologia, 7Department of Nutrition and Bioprogramming,Instituto Nacional de Perinatologia, 8Department of Human Genetics and Genomics,Instituto Nacional de Perinatologia, Mexico City, Mexico

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

Automatic Translation

Automatic Translation

该方案提供了一种高效的胶原酶消化方法，用于在单个过程中从人内脏脂肪中分离活脂肪细胞和基质血管部分-SVF细胞，包括从脂肪细胞中获得高质量RNA的方法，以及通过对多种膜结合标记物进行染色以通过流式细胞术分析的SVF-巨噬细胞表型。

Tags

AdipocytesStromal Vascular FractionRNA AnalysisMacrophage PhenotypingEnzymatic DigestionCollagenaseFlow CytometryGreater Omentum

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