Name: standardized preparation of single-cell suspensions from mouse lung tissue using the gentlemacs dissociator
Uploaded: 2009-07-02T00:00:00
Description: Watch this Scientific Journal Video about Standardized Preparation of Single-Cell Suspensions from Mouse Lung Tissue using the gentleMACS Dissociator at JoVE.com

<p class="jove_content"> De auteurs zijn medewerkers van Miltenyi Biotec GmbH, Duitsland. </p>

<p class="jove_content"> Om te werken onder steriele omstandigheden, is het raadzaam om alle stappen in een laminaire stroming kap uit te voeren. </p><p class="jove_title"> 1. Materialen </p><ol><li> HEPES buffer: 10 mM HEPES-NaOH pH 7,4, 150 mM NaCl, 5 mM KCl, 1 mM <sub>MgCl2,</sub> 1,8 mM CaCl <sub>2)</sub> </li><li> Collagenase D oplossing: Collagenase D: 100 mg / ml (Collagenase D&gt; 0,15 U / mg), in HEPES buffer </li><li> DNase I oplossing: 20.000 U / ml DNase I </li><li> PBS buffer bij pH 7,2 </li><li> PEB buffer: een deel BSA Stock Solu...

<ol>
	<li>Swanson, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flt3-Ligand,+IL-4,+GM-CSF,+and+Adherence-Mediated+Isolation+of+Murine+Lung+Dendritic+Cells:+Assessment+of+Isolation+Technique+on+Phenotype+and+Function.">Flt3-Ligand, IL-4, GM-CSF, and Adherence-Mediated Isolation of Murine Lung Dendritic Cells: Assessment of Isolation Technique on Phenotype and Function.</a> <em style='font-style: italic'>J. Immunol</em>. <b>173</b>, 4875-4881 (2004).</li><li>Vermaelen, K., Pauwels, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accurate+and+simple+discrimination+of+mouse+pulmonary+dendritic+cell+and+macrophage+populations+by+flow+cytometry:+Methodology+and+new+insights.">Accurate and simple discrimination of mouse pulmonary dendritic cell and macrophage populations by flow cytometry: Methodology and new insights.</a> <em style='font-style: italic'>Cytometry Part A</em>. <b>61A</b>, 170-177 (2004).</li></ol>

<p class="jove_content"> In deze video, introduceren we een nieuwe methode voor de dissociatie van de muis longweefsel. We laten zien dat een combinatie van mechanische en enzymatische behandeling van het longweefsel een hoog percentage van levensvatbare leukocyten en endotheelcellen opgeleverd. In het bijzonder, was de mechanische desintegratie van het weefsel bereikt door de gentleMACS Dissociator. De gentleMACS C Tubes zijn een rotor - stator systeem, dat scheidt weefsel in een zachte manier. De procedure wordt gecontroleerd door de programm...

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		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
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<tr>
<td>C Tubes</td>
<td>&nbsp;</td>
<td>Miltenyi Biotec</td>
<td>Order no. 130-093-237</td>
<td><a href="http://www.miltenyibiotec.com/en/PG_709_765_gentleMACS_C_Tubes.aspx">http://www.miltenyibiotec.com/en/PG_709_765_gentleMACS_C_Tubes.aspx</a></td>
</tr>
<tr>
<td>gentleMACS&#x2122; Starting Kit</td>
<td>&nbsp;</td>
<td>Miltenyi Biotec</td>
<td>Order no. 130-093-235</td>
<td><a href="http://www.miltenyibiotec.com/en/PG_709_763_gentleMACS_Dissociator.aspx">http://www.miltenyibiotec.com/en/PG_709_763_gentleMACS_Dissociator.aspx</a></td>
</tr>
<tr>
<td>MACSmix&#x2122; Tube Rotator</td>
<td>&nbsp;</td>
<td>Miltenyi Biotec</td>
<td>Order no. 130-090-753</td>
<td><a href="http://www.miltenyibiotec.com/en/PG_34_251_MACSmix_Tube_Rotator.aspx">http://www.miltenyibiotec.com/en/PG_34_251_MACSmix_Tube_Rotator.aspx</a></td>
</tr>
<tr>
<td>autoMACS&#x2122; Rinsing Solution</td>
<td>&nbsp;</td>
<td>Miltenyi Biotec</td>
<td>Order no. 130-091-222</td>
<td><a href="http://www.miltenyibiotec.com/en/PG_134_155_autoMACS_Rinsing_Solution.aspx">http://www.miltenyibiotec.com/en/PG_134_155_autoMACS_Rinsing_Solution.aspx</a></td>
</tr>
<tr>
<td>MACS&#174; BSA Stock Solution</td>
<td>&nbsp;</td>
<td>Miltenyi Biotec</td>
<td>Order no. 130-091-376</td>
<td><a href="http://www.miltenyibiotec.com/en/PG_33_295_MACS_BSA_Stock_Solution.aspx">http://www.miltenyibiotec.com/en/PG_33_295_MACS_BSA_Stock_Solution.aspx</a></td>
</tr>
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<td>CD11c MicroBeads</td>
<td>&nbsp;</td>
<td>Miltenyi Biotec</td>
<td>Order No. 130-052-001</td>
<td><a href="http://www.miltenyibiotec.com/en/PG_93_132_CD11c_MicroBeads.aspx">http://www.miltenyibiotec.com/en/PG_93_132_CD11c_MicroBeads.aspx</a></td>
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<td>CD11c-FITC, CD11c-PE</td>
<td>&nbsp;</td>
<td>Miltenyi Biotec</td>
<td>Order no. 130-092-026	130-091-830</td>
<td><a href="http://www.miltenyibiotec.com/en/PG_599_329_CD11c.aspx">http://www.miltenyibiotec.com/en/PG_599_329_CD11c.aspx</a></td>
</tr>
<tr>
<td>Anti-mPDCA-1-APC</td>
<td>&nbsp;</td>
<td>Miltenyi Biotec</td>
<td>Order no. 130-091-963</td>
<td><a href="http://www.miltenyibiotec.com/en/PG_599_325_Anti_mPDCA_1.aspx">http://www.miltenyibiotec.com/en/PG_599_325_Anti_mPDCA_1.aspx</a></td>
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<td>CD146-FITC</td>
<td>&nbsp;</td>
<td>Miltenyi Biotec</td>
<td>Order no. 130-092-026</td>
<td><a href="http://www.miltenyibiotec.com/en/PG_599_417_CD146_LSEC_.aspx">http://www.miltenyibiotec.com/en/PG_599_417_CD146_LSEC_.aspx</a></td>
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<td>CD45-PE</td>
<td>&nbsp;</td>
<td>Miltenyi Biotec</td>
<td>Order no. 130-091-610</td>
<td><a href="http://www.miltenyibiotec.com/en/PG_599_331_CD45.aspx">http://www.miltenyibiotec.com/en/PG_599_331_CD45.aspx</a></td>
</tr>
<tr>
<td>Propidium Iodide Solution</td>
<td>&nbsp;</td>
<td>Miltenyi Biotec</td>
<td>Order no. 130-093-233</td>
<td><a href="http://www.miltenyibiotec.com/en/PG_337_744_Propidium_Iodide_Solution.aspx">http://www.miltenyibiotec.com/en/PG_337_744_Propidium_Iodide_Solution.aspx</a></td>
</tr>
<tr>
<td>Art 1000 Reach Pipet Tips</td>
<td>&nbsp;</td>
<td>Molecular BioProducts</td>
<td>Order no. 2079</td>
<td><a href="http://www.mbpinc.com/ASP/products.aspx?query_TipID=61">http://www.mbpinc.com/ASP/products.aspx?query_TipID=61</a><br /><a href="http://www.mbpinc.com/html/products/art/display.html">http://www.mbpinc.com/html/products/art/display.html</a>
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standardized preparation of single-cell suspensions from mouse lung tissue using the gentlemacs dissociator

<p class="jove_content">De voorbereiding van single-cell schorsingen van weefsel is een belangrijke voorwaarde voor veel experimenten in cellulair onderzoek. Het proces van dissociëren hele organen vereist specifieke parameters om een ​​groot aantal levensvatbare cellen te verkrijgen op een reproduceerbare manier. De gentleMACS Dissociator optimaliseert deze taak met een eenvoudige, praktische protocol. Het instrument bevat voorgeprogrammeerde instellingen die zijn geoptimaliseerd voor het efficiënt, maar zachte dissociatie van een verscheidenheid van weefsels types, inclusief muis longen. In deze publicatie het gebruik van de gentleMACS Dissociator op longweefsel zijn afgeleid van muizen is aangetoond.</p>

Watch this Scientific Journal Video about Standardized Preparation of Single-Cell Suspensions from Mouse Lung Tissue using the gentleMACS Dissociator at JoVE.com

Standardized Preparation of Single-Cell Suspensions from Mouse Lung Tissue using the gentleMACS Dissociator

<p class="jove_content">Dissociëren cellen van specifiek weefsel soorten vereist specifieke parameters voor weefsel opwinding tot het verkrijgen van een hoog volume van levensvatbare, kweekbare cellen. De Miltenyi gentleMACS dissociator optimaliseert deze taak met een eenvoudige, praktische protocol. In deze publicatie het gebruik van dit apparaat op longweefsel wordt uitgelegd.</p>

Gestandaardiseerde Voorbereiding van de Single-celsuspensies van Mouse longweefsel met behulp van de gentleMACS Dissociator

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