Preparation of a Single-Cell Suspension from Mouse Carotid Arteries for Single-Cell Sequencing

作者： Fengchan Li1, Zhen Zhu1, Yun Du1, Li Zhu1,2,3, Chaojun Tang1,2,3 1Cyrus Tang Hematology Center, Cyrus Tang Medical Institute,Soochow University, 2Collaborative Innovation Center of Hematology of Jiangsu Province,Soochow University, 3Suzhou Key Laboratory of Thrombosis and Vascular Biology,Soochow University

简介：

Overview

This study addresses the challenges of low cellular viability and large cell debris in vascular tissue dissociation by presenting a two-step cell digestion protocol specifically for mouse carotid arteries. The protocol yields a high viability single-cell suspension that is adaptable for other vessels, advancing techniques in cardiovascular research.

Key Study Components

Research Area

• Cardiovascular research
• Cell dissociation methods
• Single-cell analysis

Background

• Common issues with vascular tissue dissociation include low cellular viability and large debris.
• Effective dissociation methods are crucial for studying various cell types within blood vessels.
• Previous protocols may lack adaptability or require specialized equipment.

Methods Used

• Two-step enzymatic cell digestion
• Mouse carotid arteries
• Filtration and centrifugation techniques

Main Results

• 176,000 single cells were successfully collected from mouse carotid arteries.
• The protocol enhanced dissociation efficiency and cell viability.
• Unsupervised clustering identified four main cell types in the carotid arteries.

Conclusions

• The study provides an effective protocol for preparing single-cell suspensions from carotid arteries.
• This methodology has implications for targeted therapies in cardiovascular diseases.

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