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All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Animals
<ol>
	<li>Maintain adult 3- to 6-week-old female mice in a room with controlled temperature and humidity (with phases of 12L:12D and fed&#160;ad libitum).</li>
	<li>Inject mice intraperitoneally with 2.5 IU of Pregnant mare serum gonadotropin (PMSG) to stimulate follicular growth by mimicking the effects of folli...

mouse antral oocyte isolation: a method to isolate antral oocytes from freshly harvested mouse ovaries

Source: Monti, M. et al. <a href="https://www.jove.com/video/53616" target="_blank">Isolation and Characterization of Mouse Antral Oocytes Based on Nucleolar Chromatin Organization</a>. J. Vis. Exp. (2016)
This video describes the isolation of antral oocytes from freshly harvested mouse ovaries. These antral oocytes can be used for further characterization and developmental studies.

<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/20757/20757_Figure1.jpg" /> 
Figure 1. Preparation of a mouth pipette using glass Pasteur pipettes and a Bunsen burner.
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/20757/20757_Figure2.jpg" /> 
Figure 2. Intact ovaries in Petri dish filled with pre-warmed and equilibrated M2 medium.</stron...

Mouse Antral Oocyte Isolation: A Method to Isolate Antral Oocytes from Freshly Harvested Mouse Ovaries

Source: Monti, M. et al. Isolation and Characterization of Mouse Antral Oocytes Based on Nucleolar Chromatin Organization. J. Vis. Exp. (2016)
This video ...

In a mammalian ovary, fully grown oocytes derived from the antral follicles only need a short period of maturation in culture, a crucial factor for in vitro embryogenesis. These oocytes are surrounded by a cluster of somatic cells, which must be removed through denudation to facilitate sperm injection.
To isolate denuded mouse antral oocytes, begin with an anesthetized female mouse pre-injected with pregnant mare serum gonadotropin hormone to stimulate follicular growth. Prep the mouse and incise the lower abdomen.
Exteriorize the intestine to reveal ovaries at the top of V-shaped uterine tubes. Dissect an ovary. Transfer it to a pre-equilibrated culture medium to maintain the viability of oocytes. Under a stereomicroscope, identify the antral follicles by their distinctive fluid-filled cavity.
Using a sterile needle, puncture the antral follicles of the ovary to release oocytes. Using a pipette of an optimum diameter, carefully aspirate the oocytes to protect them from mechanical damage. Avoid aspirating follicular cells and other tissue debris.
Subsequently, transfer the oocytes into multiple media drops covered with mineral oil. This step helps remove the surrounding somatic cells and generate denuded oocytes. Lastly, transfer the denuded oocytes into fresh media drops with mineral oil and preserve them for further experiments.

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This video describes the isolation of antral oocytes from freshly harvested mouse ovaries. These antral oocytes can be used for further characterization and developmental studies.

Overview

Protocol

<img title="figure-representative results-58" alt="figure-representative results-58" class="xfigimg" src="/files/ftp_upload/20757/20757_Figure1.jpg" /> 
Figure 1. Preparation of a mouth pipette using glass Pasteur pipettes and a Bunsen burner.
<img title="figure-representative results-337" alt="figure-representative results-337" class="xfigimg" src="/files/ftp_upload/20757/20757_Figure2.jpg" /> 
Figure 2. Intact ovaries in Petri dish filled with pre-warmed and equilibrated M2 medium.</stron...

Mouse Antral Oocyte Isolation: A Method to Isolate Antral Oocytes from Freshly Harvested Mouse Ovaries

In This Article

Overview

Protocol

Representative Results

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