Journal
/
Medicine
/
Isolation of Viable Adipocytes and Stromal Vascular Fraction from Human Visceral Adipose Tissue Suitable for RNA Analysis and Macrophage Phenotyping
JoVE Journal
Medicine
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Medicine
Isolation of Viable Adipocytes and Stromal Vascular Fraction from Human Visceral Adipose Tissue Suitable for RNA Analysis and Macrophage Phenotyping

Isolation of Viable Adipocytes and Stromal Vascular Fraction from Human Visceral Adipose Tissue Suitable for RNA Analysis and Macrophage Phenotyping

DOI:
10.3791/61884-v

•

06:22 min

•

October 27, 2020

•

, , , , , , , ,
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

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.

Tags

Keywords: AdipocytesStromal Vascular FractionRNA AnalysisMacrophage PhenotypingEnzymatic DigestionCollagenaseFlow CytometryGreater Omentum

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/61884?article_type=v

Isolation of Human Islets from Partially Pancreatectomized Patients
check_url/61884?article_type=v

Experimental Generation of Carcinoma-Associated Fibroblasts (CAFs) from Human Mammary Fibroblasts
check_url/61884?article_type=v

Isolation of Cardiomyocyte Nuclei from Post-mortem Tissue
check_url/61884?article_type=v

Encapsulation Thermogenic Preadipocytes for Transplantation into Adipose Tissue Depots
check_url/61884?article_type=v

Isolation and Cellular Phenotyping of Mesenchymal Stem Cells Derived from Synovial Fluid and Bone Marrow of Minipigs
check_url/61884?article_type=v

Stromal Vascular Fraction-enriched Fat Grafting for the Treatment of Symptomatic End-neuromata
check_url/61884?article_type=v

Assessment of Human Adipose Tissue Microvascular Function Using Videomicroscopy
check_url/61884?article_type=v

Isolation of Atrial Cardiomyocytes from a Rat Model of Metabolic Syndrome-related Heart Failure with Preserved Ejection Fraction
check_url/61884?article_type=v

Clinical Protocol of Producing Adipose Tissue-Derived Stromal Vascular Fraction for Potential Cartilage Regeneration
check_url/61884?article_type=v

Isolation of Adipose Derived Regenerative Cells for the Treatment of Erectile Dysfunction Following Radical Prostatectomy

Read Article