Isolation of Enteric Glial Cells from the Submucosa and Lamina Propria of the Adult Mouse

Zhen Wang; Ramon Ocadiz-Ruiz; Sinju Sundaresan; Lin Ding; Michael Hayes; Nirakar Sahoo; Haoxing Xu; Juanita L. Merchant

doi:10.3791/57629

Isolation of Enteric Glial Cells from the Submucosa and Lamina Propria of the Adult Mouse

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Isolation of Enteric Glial Cells from the Submucosa and Lamina Propria of the Adult Mouse

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07:53 min

August 15, 2018

DOI: 10.3791/57629-v

Zhen Wang^1,4, Ramon Ocadiz-Ruiz¹, Sinju Sundaresan¹, Lin Ding¹, Michael Hayes¹, Nirakar Sahoo³, Haoxing Xu^1,2, Juanita L. Merchant^1,2,5

¹Department of Internal Medicine-Gastroenterology,University of Michigan, ²Department of Molecular and Integrative Physiology,University of Michigan, ³Department of Molecular, Cellular and Developmental Biology,University of Michigan, ⁴Department of Gastrointestinal Surgery,The First Affiliated Hospital of Guangxi Medical University, ⁵Division of Gastroenterology,University of Arizona College of Medicine

Overview

This article describes a rapid, non-enzymatic method for isolating enteric-glial cells from the intestinal submucosa. The technique utilizes sequential EDTA incubations to chelate divalent cations, followed by incubation in a non-enzymatic cell recovery solution, resulting in a highly enriched culture for functional analysis.

Key Study Components

Area of Science

Neuroscience
Cell Biology
Enteric Nervous System

Background

Enteric-glial cells play a crucial role in the enteric nervous system.
Isolation of these cells is essential for studying their function.
Traditional methods may involve enzymatic treatments, which can be time-consuming.
This study presents a non-enzymatic approach to enhance efficiency.

Purpose of Study

To develop a rapid method for isolating enteric-glial cells.
To facilitate functional analysis of these cells.
To improve understanding of the enteric nervous system's role in gut physiology.

Methods Used

Sequential EDTA incubations to chelate divalent cations.
Non-enzymatic cell recovery solution for cell isolation.
Plating on poly-D-lysine and laminin for cell culture.
Dissection techniques to obtain intestinal samples.

Main Results

Successful isolation of enteric-glial cells from the intestinal submucosa.
High enrichment of submucosal glial cells for analysis.
Demonstrated efficiency of the non-enzymatic method.
Potential for further research into the enteric nervous system.

Conclusions

The non-enzymatic method is effective for isolating enteric-glial cells.
This technique can advance research in neuroscience.
Further studies can explore the functional roles of these cells.

Frequently Asked Questions

What are enteric-glial cells?

Enteric-glial cells are supportive cells in the enteric nervous system that play a role in gut function.

Why is a non-enzymatic method preferred?

Non-enzymatic methods are faster and may preserve cell functionality better than enzymatic methods.

What is the significance of isolating these cells?

Isolating enteric-glial cells allows researchers to study their specific functions and roles in gut health.

How does this method compare to traditional techniques?

This method is quicker and avoids the potential damage caused by enzymatic treatments.

What applications can arise from this research?

The findings can lead to better understanding of gut disorders and the enteric nervous system's role in health.

Can this method be applied to other cell types?

While this method is tailored for enteric-glial cells, similar techniques may be adapted for other cell types.

Here, we describe the isolation of enteric-glial cells from the intestinal-submucosa using sequential EDTA incubations to chelate divalent cations and then incubation in non-enzymatic cell recovery solution. Plating the resultant cell suspension on poly-D-lysine and laminin results in a highly enriched culture of submucosal glial cells for functional analysis.

This method can help answer key questions in the neuroscience field, specifically the function of the enteric nervous system. The main advantage of the technique is that it is a rapid, non-enzymatic method to isolate enteric cells from the lamina propria and submucosa rather than the smooth muscle. Begin by identifying the distal stomach and pylorus.

Hold the pylorus with forceps while snipping away the mesentery. Then use blunt dissection with the back of the scissors to scrape away adherent pancreas. Then use scissors to remove seven centimeters of the proximal intestine without tearing.

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