Isolation, Culture, and Adipogenic Induction of Stromal Vascular Fraction-derived Preadipocytes from Mouse Periaortic Adipose Tissue

Min Liang; Yijie Huang; Yangjing Jiang; Yunwen Hu; Zhaohua Cai; Ben He

doi:10.3791/65703

Isolation, Culture, and Adipogenic Induction of Stromal Vascular Fraction-derived Preadipocytes f...

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Isolation, Culture, and Adipogenic Induction of Stromal Vascular Fraction-derived Preadipocytes from Mouse Periaortic Adipose Tissue

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06:56 min

July 21, 2023

DOI: 10.3791/65703-v

Min Liang*¹, Yijie Huang*¹, Yangjing Jiang¹, Yunwen Hu¹, Zhaohua Cai¹, Ben He¹

¹Department of Cardiology, Shanghai Chest Hospital,Shanghai Jiao Tong University School of Medicine

Overview

This study addresses the standardized isolation, culture, and adipogenic induction of stromal vascular fraction-derived preadipocytes from mouse periaortic adipose tissue (PVAT). The findings provide insights into PVAT function and its interactions with vascular cells, paving the way for further exploration of adipogenesis and cellular communication.

Key Study Components

Research Area

Preadipocyte isolation and culture
Adipogenic induction
Perivascular adipose tissue (PVAT) function

Background

Challenges in PVAT availability and resource requirements
Significance of perivascular adipose tissue in vascular biology
Need for in vitro studies with primary preadipocytes

Methods Used

In vivo dissection and isolation techniques from mouse PVAT
Cell culture and adipogenic induction protocols
Microsurgery and enzymatic digestion methods

Main Results

Successful isolation of stromal vascular fraction from PVAT
Establishment of a robust culture system for preadipocytes
Framework for studying adipocyte-vascular cell interactions

Conclusions

This study provides a reliable method for isolating preadipocytes, facilitating research on PVAT.
The findings highlight the intricate relations between preadipocytes and vascular cells, contributing to greater understanding in biology research.

Frequently Asked Questions

What is perivascular adipose tissue?

Perivascular adipose tissue (PVAT) is the fat tissue surrounding blood vessels, playing a role in vascular function.

Why is the isolation of preadipocytes important?

Isolating preadipocytes allows researchers to study adipogenesis and interactions with other cell types in a controlled environment.

What are the challenges faced in isolating PVAT?

Challenges include limited availability of PVAT and the resource demands compared to using established cell lines.

How does adipogenic induction work?

Adipogenic induction involves stimulating preadipocytes to develop into mature adipocytes through specific growth factors and environmental conditions.

What can be studied using this isolation method?

This method enables studies on adipogenesis, the function of PVAT, and the signaling pathways involved in fat cell differentiation.

Are there any specific technologies used in this research?

Yes, technologies such as microscopy and enzymatic assays are utilized to analyze cells and their functions.

Here, we describe the isolation, culture, and adipogenic induction of stromal vascular fraction-derived preadipocytes from mouse periaortic adipose tissue, allowing for the study of perivascular adipose tissue function and its relationship with vascular cells.

We aim to establish a standardized approach for the isolation, culture, and adipogenic induction of stromal vascular fraction-derived preadipocytes from mass PVAT. This protocol enables us to conduct further investigations into PVAT function and into the communication between periaortic adipocytes and vascular cells in vitro. The main challenges we currently face in our experiments include the restricted of availability of PVAT and the requirements for additional human and the material resources when compared to using immortalized preadipocytes.

Our protocol allows for the study of perivascular adipose tissue function and it's relationship with vascular cells. We also offers a basis for studying PVAT ring and genes, and heterogeneity, and provides a platform for exploring the important regulator of PVAT differentiation and the adipogenesis in vitro. Begin by spraying the properly euthanized mouse with 75%alcohol to disinfect the skin.

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