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使用组织解离器从小鼠白色脂肪组织中半自动分离基质血管部分
使用组织解离器从小鼠白色脂肪组织中半自动分离基质血管部分
JoVE Journal
Biology
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JoVE Journal Biology
Semi-Automated Isolation of the Stromal Vascular Fraction from Murine White Adipose Tissue Using a Tissue Dissociator

使用组织解离器从小鼠白色脂肪组织中半自动分离基质血管部分

Full Text
3,005 Views
06:08 min
May 19, 2023

DOI: 10.3791/65265-v

Kaede Saito1,2, Yuta Hiraike1,2,3, Misato Oguchi2, Toshimasa Yamauchi2

1Division for Health Service Promotion,The University of Tokyo, 2Department of Diabetes and Metabolic Diseases, Graduate School of Medicine,The University of Tokyo, 3The University of Tokyo Excellent Young Researcher Program,The University of Tokyo

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This study focuses on the semi-automated isolation of the stromal vascular fraction (SVF) from murine adipose tissue, with the aim of obtaining preadipocytes and promoting adipocyte differentiation in vitro. A novel protocol utilizing a tissue dissociator for collagenase digestion is presented, which enhances reproducibility and reduces experimental variation.

Key Study Components

Research Area

  • Adipocyte differentiation
  • Energy expenditure in obesity treatments
  • Murine models for metabolic research

Background

  • Different functions of brown, beige, and white adipocytes
  • Need for reproducible methods in adipocyte research
  • Identification of critical transcription factors regulating differentiation

Methods Used

  • Semi-automated isolation using collagenase digestion
  • Murine adipose tissue
  • Tissue dissociators and enzymatic assays

Main Results

  • Improved methods for isolating SVF and differentiating adipocytes
  • Successful differentiation demonstrated via Oil Red O staining and mRNA analysis
  • Investigation into the effects of Rosiglitazone on brown fat-specific gene expression

Conclusions

  • This protocol facilitates reliable SVF isolation for adipocyte differentiation.
  • The findings hold significant implications for obesity and diabetes research.

Frequently Asked Questions

What is the significance of the stromal vascular fraction in adipocyte research?
The SVF is crucial for isolating preadipocytes, which are necessary for studying adipocyte development and differentiation.
How does the tissue dissociator improve the isolation process?
It reduces variability and contamination during collagenase digestion, leading to more reproducible results.
What specific outcomes were measured to confirm adipocyte differentiation?
Outcomes included Oil Red O staining and mRNA expression levels of adipogenesis regulators.
What are the potential applications of the differentiated adipocytes?
They can be used for functional and mechanistic analyses, including oxygen consumption rate analysis.
How might this research contribute to obesity treatments?
By promoting energy expenditure rather than suppressing intake, new obesity treatments can be developed based on these findings.
What role does the transcription factor NFIA play in this study?
NFIA is identified as a critical regulator of adipocyte differentiation and metabolism.

该协议描述了从小鼠脂肪组织中半自动分离基质血管部分(SVF)以获得前脂肪细胞并在 体外实现脂肪细胞分化。使用组织解离剂进行胶原酶消化可减少实验变异并提高可重复性。

我们对脂肪细胞的棕色和米色亚型感兴趣,它们以热量的形式耗散能量,而白色脂肪细胞通常储存能量 我们的目标是开发一种基于促进能量消耗而不是抑制能量摄入的肥胖和糖尿病的新疗法。我们已经通过全基因组开放染色质分析确定了转录因子NFI作为棕色脂肪细胞分化的关键调节因子。NFI和脂肪生成的主要转录调节因子PPAR-γ共同定位为棕色脂肪特异性增强子。

在从小鼠脂肪组织中分离基质血管部分的过程中,手动切碎和消化脂肪组织的胶原酶会导致实验变异性和污染。我们的方法使用组织解离剂进行胶原酶消化,这有助于SVF分离,减少实验变异性和污染,并提高可重复性。使用个体分化的脂肪细胞进行实验对于我们领域的研究是必不可少的。

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