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Biology
Aislamiento de células endoteliales de la luz de arterias carótidas de ratón para experimentos mu...
Aislamiento de células endoteliales de la luz de arterias carótidas de ratón para experimentos mu...
JoVE Journal
Biology
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JoVE Journal Biology
Isolation of Endothelial Cells from the Lumen of Mouse Carotid Arteries for Single-Cell Multi-Omics Experiments

Aislamiento de células endoteliales de la luz de arterias carótidas de ratón para experimentos multiómicos unicelulares

Full Text
3,893 Views
09:12 min
October 4, 2021

DOI: 10.3791/63128-v

Sandeep Kumar*1, Aitor Andueza*1, Nicolas Villa-Roel1, Juyoung Kim1, Dong-Won Kang1, Hanjoong Jo1,2

1Wallace H. Coulter Department of Biomedical Engineering,Georgia Institute of Technology and Emory University, 2Division of Cardiology,Georgia Institute of Technology and Emory University

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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 presents a novel protocol for isolating endothelial cells and nuclei from mouse carotid arteries under various flow conditions, enabling detailed examination of molecular pathways related to atherosclerosis. The method enhances the ability to perform single-cell omics experiments, facilitating the study of endothelial cell responses to disturbed blood flow.

Key Study Components

Research Area

  • Single-cell omics
  • Endothelial cell biology
  • Atherosclerosis research

Background

  • Endothelial cells are difficult to study due to their low abundance in blood vessels.
  • The protocol significantly improves the yield of endothelial enriched samples.
  • Understanding molecular pathways influenced by blood flow dynamics is crucial for therapeutic development.

Methods Used

  • Isolation of endothelial cells and nuclei from carotid arteries of mice.
  • In vivo experimentation with various flow conditions.
  • Utilization of enzymatic digestion and subsequent cell processing techniques.

Main Results

  • Successful extraction and analysis of single endothelial cells for gene expression profiling.
  • Identification of responses to both acute and chronic disturbed flow conditions.
  • Establishment of potential therapeutic targets for atherosclerosis.

Conclusions

  • This method demonstrates a robust approach for studying endothelial cells in a single-cell context.
  • Findings have significant implications for understanding vascular biology and developing treatments for vascular diseases.

Frequently Asked Questions

What is the significance of isolating endothelial cells?
Isolating endothelial cells allows for targeted studies of their functions and responses in various physiological and pathological environments.
How does disturbed flow affect endothelial cells?
Disturbed blood flow can lead to changes in gene expression that contribute to the development of vascular diseases, such as atherosclerosis.
What techniques are used in this study?
The study employs enzymatic digestion, centrifugation, and single-cell sequencing techniques to analyze isolated cells.
Can this method be applied to other vascular tissues?
Yes, the method can potentially be adapted to isolate endothelial cells from other blood vessels.
What are potential applications of this research?
The research can inform the development of new therapies targeting endothelial function in various cardiovascular conditions.
What are some challenges in carotid artery isolation?
Challenges include the meticulous dissection required to avoid damage and contamination during the isolation process.
What is the role of flow conditions in vascular biology?
Flow conditions are crucial in determining endothelial cell behavior, which influences vascular health and disease.

Presentamos un método para aislar células endoteliales y núcleos de la luz de arterias carótidas de ratón expuestas a condiciones de flujo estables o alteradas para realizar experimentos ómicos unicelulares.

En un vaso sanguíneo, el número de células endoteliales es muy bajo, lo que dificulta su estudio. Nuestro protocolo elimina esta limitación y nos permite estudiar nuevas vías moleculares y terapéuticas. Nuestro protocolo nos permite obtener muestras enriquecidas endotelialmente, y estudiar cómo el flujo perturbado agudo y crónico afecta a este tipo celular.

La capacidad de estudiar las células endoteliales con más detalle permite definir genes diana y, por lo tanto, terapias potenciales para la aterosclerosis dirigidas a las células endoteliales. El aislamiento de la arteria carótida es complicado y requiere paciencia y experiencia. Para comenzar, prepare al animal inyectando 0.05 miligramos por kilogramo de buprenorfina por vía subcutánea.

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