<p class="jove_content">I would like to thank Mike Dewey from the Peromyscus Genetic Stock Center for his help, patience and knowledge.</p>

<p class="jove_title">Embryo/Oocyte retrieval</p>
<ol>
<li>At least two hours prior to retrieval of embryos/oocytes, KSOM is placed in the 37&ordm;C CO<sub>2</sub> incubator and FHM must be thawed from -20&deg;. A glass dish containing three concave shallow wells is used to house drops of KSOM in the incubator. To prevent dehydration, the KSOM and dish are placed in a lidded container that has water at the bottom. This apparatus prevents evaporation without the use of mineral oil.<br /><br /></li>
<li>The oviducts from an ovulating female are p...

<p class="jove_content">In this article, we demonstrate the retrieval of oocytes from ovulation-induced (superovulated) Peromyscus.  The use of superovulation (as opposed to natural estrous cycle) is common due to the much greater resulting numbers.  However, caution must be taken in interpreting results utilizing such oocytes/embryos, as both the hormones used to induce ovulation and culturing may have effects.</p>

<p class="jove_content">When estrous has been confirmed, females are euthenized and oviducts recovered along with a small porti...

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		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Flushing needle</td>
<td>Tool</td>
<td>Zephyrtronics</td>
<td>ZT5-130-L</td>
<td>30 gauge blunt</td>
</tr>
<tr>
<td>Forceps (2)</td>
<td>Tool</td>
<td>Roboz</td>
<td>RS-5060</td>
<td>&nbsp;</td>
<tr>
<td>Peromyscus</td>
<td>Animal</td>
<td>Peromyscus Genetic Stock Center</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Scissors</td>
<td>Tool</td>
<td>Roboz</td>
<td>RS-5880</td>
<td>&nbsp;</td>
<tr>
<td>FHM HEPES Buffered Media</td>
<td>Reagent</td>
<td>Specialty Media</td>
<td>MR-025-d</td>
<td>&nbsp;</td>
<tr>
<td>KSOM Embryo Culture Medium (5x10 ml)</td>
<td>Reagent</td>
<td>Specialty Media</td>
<td>MR-020P-5F</td>
<td>&nbsp;</td>
<tr>
<td>Mouth Pipette</td>
<td>Tool</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Pulled from glass capillaries and attached to rubber tubing</td>
</tr>
<tr>
<td>Oocyte holding dish</td>
<td>Tool</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Glass container with concave divets</td>
</tr>		</tbody>
		</table>
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retrieval of mouse oocytes

To date, only a few studies have reported successful manipulations of Peromyscus embryogenesis or reproductive biology.  Together with the Peromyscus Genetic Stock Center (http://stkctr.biol.sc.edu), we are characterizing the salient differences needed to develop this system.  A primary goal has been to optimize oocyte/early embryo retrieval.

Watch this Scientific Journal Video about Retrieval of Mouse Oocytes at JoVE.com

Retrieval of Mouse Oocytes

<p class="jove_content">This protocol illustrates the technique for extracting oocytes or early-stage fertilized embryos from the oviduct of mice. The ability to identify the infindibulum and insert a blunt end needle into it is essential to correctly performing the procedure.</p>

To date, only a few studies have reported successful manipulations of Peromyscus embryogenesis or reproductive biology.  Together with the Peromyscus ...

Research

JoVE Journal

Biology

27.1K vues.  University of California, Irvine (UCI). Ce protocole illustre la technique d'extraction ou d'ovocytes à un stade précoce des embryons fertilisés de l'oviducte de la souris. La capacité à identifier les infindibulum et insérer une aiguille extrémité émoussée en elle est essentielle à l'exécution correcte de la procédure.