Name: preparation of dissociated mouse cortical neuron cultures
Uploaded: 2007-12-19T00:00:00
Description: Watch this Scientific Journal Video about Preparation of Dissociated Mouse Cortical Neuron Cultures at JoVE.com

<p class="jove_step"> Voorbereidingen voor de dag van het kweken: </p><ul><li> Bereid steriele ontleden oplossing (DS). </li><li> Bereid NBM/B27 (Neurobasal Mediom met B27 supplementen). </li><li> Autoclaaf DDH <sub>2</sub> O en steriliseren glas coversilps, indien nodig. </li><li> Vacht weefselkweek gerechten of glazen dekglaasjes met poly-D-lysine. </li></ul><p class="jove_step"> Poly-D-Lysine Coating: </p><p class="jove_content"> Bereid de dag voor het kweken van onder steriele omstandigheden. </p><ul><li> Dooi hoeveelheid van 10X PDL en leg...

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		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>5-Fluoro-2'-deoxyuridine</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>F0503</td>
<td>&nbsp;</td>
<tr>
<td>Sodium Chloride</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>S9625</td>
<td>&nbsp;</td>
<tr>
<td>Vibraslicer</td>
<td>&nbsp;</td>
<td>Campden Instruments Ltd.</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Potassim Chloride</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>P4504</td>
<td>&nbsp;</td>
<tr>
<td>Sodium Phosphate Dibasic</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>S0876</td>
<td>&nbsp;</td>
<tr>
<td>Potassium Phosphate Monobasic</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>P5379</td>
<td>&nbsp;</td>
<tr>
<td>Hepes</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>H3375</td>
<td>&nbsp;</td>
<tr>
<td>D (+)-Glucose</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>G8270</td>
<td>&nbsp;</td>
<tr>
<td>Sucrose</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>S0389</td>
<td>&nbsp;</td>
<tr>
<td>Poly-D-Lysine</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>P7280</td>
<td>&nbsp;</td>
<tr>
<td>B27 Supplement</td>
<td>&nbsp;</td>
<td>Invitrogen (Gibco)</td>
<td>17504-044</td>
<td>&nbsp;</td>
<tr>
<td>Neurobasal Media (NBM)</td>
<td>&nbsp;</td>
<td>Invirtogen (Gibco)</td>
<td>21103-049</td>
<td>&nbsp;</td>
<tr>
<td>2-Amino-5-phosphonopentanoic acid</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>A5282</td>
<td>&nbsp;</td>
<tr>
<td>Bovine Albumin</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>A7030</td>
<td>&nbsp;</td>
<tr>
<td>Trypsin Inhibitor</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>T9253</td>
<td>&nbsp;</td>
<tr>
<td>Sodium Hydroxide, 1N solution</td>
<td>&nbsp;</td>
<td>Fisher</td>
<td>SS266-1</td>
<td>&nbsp;</td>
<tr>
<td>L-Cysteine</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>C7755</td>
<td>&nbsp;</td>
<tr>
<td>Bacto Agar</td>
<td>&nbsp;</td>
<td>Difco</td>
<td>0140-01</td>
<td>&nbsp;</td>
<tr>
<td>Papain</td>
<td>&nbsp;</td>
<td>Worthington </td>
<td>LS 03126</td>
<td>&nbsp;</td>
<tr>
<td>Sterile 0.2 &micro;m Syringe Filter</td>
<td>&nbsp;</td>
<td>Fisher</td>
<td>DDA02025S0</td>
<td>&nbsp;</td>
<tr>
<td>Glass Coverslips, No1, &#216;12mm</td>
<td>&nbsp;</td>
<td>Bellco Glass Inc.</td>
<td>1943-00012</td>
<td>&nbsp;</td>
<tr>
<td>Minimum Essential Media (MEM)</td>
<td>&nbsp;</td>
<td>Invitrogen (Gibco)</td>
<td>11090-081</td>
<td>with Earle's salts
without L-glutamine, needed for growing non-neuronal cultures</td>
</tr>
<tr>
<td>Penicillin/Streptomycin</td>
<td>&nbsp;</td>
<td>Invitrogen (Gibco)</td>
<td>15070-063</td>
<td>for non-neuronal culture</td>
</tr>
<tr>
<td>Fetal Bovine Serum</td>
<td>&nbsp;</td>
<td>Invitrogen (Gibco)</td>
<td>16140-071</td>
<td>needed for non-neuronal cultures</td>
</tr>
<tr>
<td>Nunclon "Triple Flask" Tissue Culture Bottle</td>
<td>&nbsp;</td>
<td>Fisher</td>
<td>12-565-25</td>
<td>Use for non-neuronal cultures. these flasks are very convenient when producing "conditioned Neurobasal Media/B27", but any other tissue culture flasks or dishes can be used instead.</td>
</tr>
<tr>
<td>Glass bottom "Imaging dishes"</td>
<td>&nbsp;</td>
<td>MatTek Corp.</td>
<td>P35G-1.5-10-C</td>
<td>Glass bottomed, &#216;35mm culture dishes ideal for Calcium or Sodium Imaging when using an inverted imaging setup. But expensive!</td>
</tr>		</tbody>
		</table>
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preparation of dissociated mouse cortical neuron cultures

<p class="jove_content">Deze video zal u door het proces voor het genereren van corticale neuronale culturen uit de late embryo en de vroege postnatale hersenen van muizen. Deze culturen kunnen worden gebruikt voor een verscheidenheid aan toepassingen, waaronder immunocytochemie, biochemie, elektrofysiologie, calcium en natrium beeldvorming, eiwit en / of RNA isolatie. Deze culturen ook een platform om de neuronale ontwikkeling van transgene dieren dat een te late embryonale of postnatale dodelijk genmutatie dragen bestuderen. De procedure is relatief eenvoudig, vergt enige ervaring in weefselkweek techniek en mag niet langer duren dan twee tot drie uur of u goed voorbereid. Een zorgvuldige scheiding van de corticale schil van de thalamo-corticale fiber-darmkanaal vermindert het aantal ongewenste niet-neuronale cellen. Ter verhoging van opbrengsten van neuronale cellen voorzichtig vermaal de stukken van de corticale weefsel na het enzym incubatie stap. Dit is noodzakelijk omdat het voorkomt onnodige schade aan cellen en vroegtijdige neuronale celdood. Aangezien deze culturen worden gehandhaafd in de afwezigheid van glia feeder cellen, ze bieden ook een extra voordeel van de groeiende culturen verrijkt met neuronen.</p>

Watch this Scientific Journal Video about Preparation of Dissociated Mouse Cortical Neuron Cultures at JoVE.com

Preparation of Dissociated Mouse Cortical Neuron Cultures

<p class="jove_content">Deze video toont een procedure voor het genereren van neuronale culturen uit de late embryo en de vroege postnatale muis cortex. Deze culturen kunnen worden gebruikt voor immunocytochemie, biochemie, elektrofysiologie, calcium en natrium beeldvorming en een platform te bieden voor de neuronale ontwikkeling van transgene dieren die een postnatale dodelijke genmutatie dragen bestuderen.</p>

De voorbereiding van gescheiden Mouse Cortical Neuron culturen

This video will guide you through the process for generating cortical neuronal cultures from late embryo and early postnatal mouse brain. These cultures ...

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