Using <em>Ex Vivo</em> Upright Droplet Cultures of Whole Fetal Organs to Study Developmental Processes during Mouse Organogenesis

Sarah J. Potter; Tony DeFalco

doi:10.3791/53262

Using Ex Vivo Upright Droplet Cultures of Whole Fetal Organs to Study Developmental Proc...

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Developmental Biology

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JoVE Journal Developmental Biology

Using Ex Vivo Upright Droplet Cultures of Whole Fetal Organs to Study Developmental Processes during Mouse Organogenesis

Using Ex Vivo Upright Droplet Cultures of Whole Fetal Organs to Study Developmental Processes during Mouse Organogenesis

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09:47 min

October 21, 2015

DOI: 10.3791/53262-v

Sarah J. Potter¹, Tony DeFalco¹

¹Division of Reproductive Sciences,Cincinnati Children's Hospital Medical Center

Overview

The ex vivo upright droplet culture is an innovative technique for studying fetal organ development. This method allows for the manipulation of fetal vascularization and organogenesis while using smaller amounts of reagents.

Key Study Components

Area of Science

Neuroscience
Developmental Biology
Cell Biology

Background

Current in vitro and in vivo techniques have limitations.
Ex vivo droplet culture provides a simpler alternative.
This method facilitates the study of complex biological processes.
Small molecule inhibitors can be tested for their effects on organ development.

Purpose of Study

To examine the effects of small molecule inhibitors on fetal organ development.
To utilize an ex vivo droplet culture technique for this examination.
To assess changes in organ architecture through immunofluorescence microscopy.

Methods Used

Opening the peritoneum of a pregnant mouse.
Removing the embryo and isolating the goad me neph complex.
Culturing the goad Meris complex in a droplet.
Using immunofluorescence microscopy with cell type specific antibodies.

Main Results

Changes in organ architecture were observed.
The effects of small molecule inhibitors were documented.
The droplet culture technique proved effective for this study.
Immunofluorescence microscopy provided valuable insights.

Conclusions

The ex vivo droplet culture is a viable alternative for studying fetal organ development.
This method allows for detailed examination of organogenesis.
Small molecule inhibitors can significantly impact fetal organ development.

Frequently Asked Questions

What is the ex vivo droplet culture technique?

It is a method used to study fetal organ development by culturing isolated organ complexes in droplets.

How does this technique compare to traditional methods?

It requires smaller amounts of reagents and allows for easier manipulation of biological processes.

What are small molecule inhibitors?

They are compounds that can interfere with specific biological pathways, used to study their effects on organ development.

What role does immunofluorescence microscopy play in this study?

It is used to visualize changes in organ architecture using specific antibodies.

Can this method be applied to other areas of research?

Yes, it can be adapted for various studies involving organ development and cellular processes.

What are the potential implications of this research?

It may lead to better understanding of fetal development and potential therapeutic targets for developmental disorders.

The ex vivo upright droplet culture is an alternative to current in vitro and in vivo experimental techniques. This protocol is easy to perform and requires smaller amounts of reagent, while permitting the ability to manipulate and study fetal vascularization, morphogenesis, and organogenesis.

The overall goal of this procedure is to examine the effects of small molecule inhibitors on fetal organ development using an ex vivo droplet culture technique. This is accomplished by first opening the peritoneum of a pregnant mouse and removing the embryo containing uterus. Next, the embryos are removed from the uterus and are freed from the yolk and amniotic sacs.

Then the goad me neph complex is dissected and isolated from the embryo. Finally, the goad Meris complex is cultured in a droplet with or without a small molecule inhibitor. Ultimately, immunofluorescence microscopy is used to show changes in organ architecture using cell type specific antibodies.

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