Cell Dissociation from the Tongue Epithelium and Mesenchyme/Connective Tissue of Embryonic-Day 12.5 and 8-Week-Old Mice

Wenxin Yu; Mohamed Ishan; Zhonghou Wang; Hong-Xiang Liu

doi:10.3791/62163

Cell Dissociation from the Tongue Epithelium and Mesenchyme/Connective Tissue of Embryonic-Day 12...

JoVE Journal
Developmental Biology

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JoVE Journal Developmental Biology

Cell Dissociation from the Tongue Epithelium and Mesenchyme/Connective Tissue of Embryonic-Day 12.5 and 8-Week-Old Mice

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06:56 min

January 21, 2021

DOI: 10.3791/62163-v

Wenxin Yu¹, Mohamed Ishan¹, Zhonghou Wang¹, Hong-Xiang Liu¹

¹Regenerative Bioscience Center; Department of Animal and Dairy Science, College of Agricultural and Environmental Sciences,University of Georgia

Overview

This protocol outlines a method for dissociating high-quality single cells from the epithelium and mesenchyme of mouse tongues. It is designed to yield healthy cells suitable for various experimental analyses.

Key Study Components

Area of Science

Cell Biology
Neuroscience
Developmental Biology

Background

High yield and quality of cells are crucial for experimental analyses.
Single cell RNA sequencing and primary stem cell cultures require healthy cells.
The mammalian tongue consists of complex tissues that can be challenging to dissociate.
This protocol addresses the need for effective cell dissociation methods.

Purpose of Study

To develop a reliable protocol for isolating single cells from mouse tongues.
To enhance the quality and yield of cells for research purposes.
To facilitate studies involving various tissue compartments of the tongue.

Methods Used

Dissection of embryos from uterine horns of pregnant mice.
Isolation of the tongue from the mandible using fine forceps and mini scissors.
Transfer of dissected tissues to Tyrode's solution for cell dissociation.
Use of a dissecting microscope for precision during dissection.

Main Results

Successful dissociation of high-quality single cells from tongue tissues.
High yield of viable cells suitable for downstream applications.
Protocol applicable to both embryonic and adult mouse tongues.
Improved methodologies for studying complex tissue structures.

Conclusions

The developed protocol is effective for isolating single cells from mouse tongues.
It provides a foundation for further research in cell biology and neuroscience.
Future studies can leverage these methods for various experimental analyses.

Frequently Asked Questions

What tissues can be dissociated using this protocol?

This protocol can dissociate cells from both the epithelium and connective tissue of the mouse tongue.

Is this protocol applicable to adult mice?

Yes, the protocol is designed for both embryonic and adult mouse tongues.

What is the purpose of using Tyrode's solution?

Tyrode's solution is used to maintain cell viability during the dissociation process.

What tools are necessary for the dissection?

Mini scissors and fine forceps are essential for precise dissection of the tongue.

How does this protocol improve cell yield?

The protocol optimizes the dissection and dissociation steps to maximize cell viability and yield.

We have developed a generalized protocol to dissociate a large quantity of high-quality single cells from the epithelium and mesenchyme/connective tissue of embryonic and adult mouse tongues.

High yield and quality of cells from complex tissues are essential to commonly used experimental analysis, such as single cell RNA sequencing and primary stem cell cultures. This protocol generates healthy cells at high yield and quality from different regions and tissue compartments of the mammalian tongue, including tongue epithelium and connective tissue. To separate the epithelium from the mesenchyme of an E 12.5 mouse tongue, transfer the euthanized pregnant female mouse to the surgical area and wet the mouse abdomen with 70%ethanol to prevent fur from getting into the operating site.

Open the abdomen using dissecting scissors to expose the uterine horns carrying the embryos, dissect the uterine horns, and transfer them to a 15 milliliters of fresh Tyrode's solution in a 100 millimeter culture dish. Under a dissecting microscope, use mini scissors and fine forceps to dissect the embryos from the uterine horns. Open the mouth cavity with the fine forceps and dissect the tongue from the mandible using mini scissors.

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