简介：
Overview
This protocol describes a method to isolate distinct subpopulations of stromal cells from murine intra-abdominal white adipose tissue using fluorescence-activated cell sorting or immunomagnetic bead separation. The technique allows for the study of functionally and molecularly unique cell types within the adipose tissue.
Key Study Components
Area of Science
- Cell biology
- Adipose tissue research
- Stromal cell isolation
Background
- Adipose tissue contains various cell types that play roles in metabolism and inflammation.
- Understanding these cell populations can provide insights into metabolic diseases.
- Isolation techniques are crucial for studying specific cell functions.
- This protocol utilizes widely available tools and reagents.
Purpose of Study
- To isolate adipogenic and fibro-inflammatory stromal cell subpopulations.
- To facilitate research on the roles of these cells in metabolic processes.
- To provide a reproducible method for researchers in the field.
Methods Used
- Fluorescence-activated cell sorting (FACS)
- Immunomagnetic bead separation
- Cell lysis and filtration techniques
- Resuspension in FBS and PBS with FC block
Main Results
- Successful isolation of distinct stromal cell populations.
- Demonstration of the procedure by a senior laboratory analyst.
- Potential applications in studying adipose tissue biology.
- Validation of the method using mouse models.
Conclusions
- The protocol provides a reliable method for isolating specific cell types from adipose tissue.
- It can enhance understanding of the cellular dynamics in metabolic diseases.
- Widespread availability of reagents makes it accessible for many researchers.
What is the main advantage of this isolation technique?
The technique utilizes widely available tools and reagents, making it accessible for researchers.
Who demonstrates the procedure in the video?
Lavanya Vishvanath, a senior laboratory research analyst from Dr. Rana Gupta's laboratory.
What types of cells can be isolated using this protocol?
Adipogenic and fibro-inflammatory stromal cell subpopulations from white adipose tissue.
How are the cells processed after isolation?
Cells are resuspended in red blood cell lysis buffer and filtered through a cell strainer.
What is the significance of studying these cell populations?
Understanding these populations can provide insights into metabolic diseases and their mechanisms.
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