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"> Författarna är anställda i Miltenyi Biotec GmbH, Tyskland. </p>

<p class="jove_content"> Att arbeta under sterila förhållanden, är det rekommenderat att utföra alla steg i ett laminärt flöde huva. </p><p class="jove_title"> 1. Material </p><ol><li> HEPES buffert: 10 mM HEPES-NaOH pH 7,4, 150 mM NaCl, 5 mm KCl, 1 mM MgCl <sub>2,</sub> 1,8 mM CaCl <sub>2)</sub> </li><li> Kollagenas D lösning: kollagenas D: 100 mg / ml (kollagenas D&gt; 0,15 U / mg), i HEPES buffert </li><li> DNas Jag lösning: 20.000 U / mL DNas jag </li><li> PBS-buffert vid pH 7,2 </li><li> PEB buffert: 1 del BSA stamlösning i 20 del...

<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"> I denna video, introducerar vi en ny metod för dissociation av musen lungvävnad. Vi visar att en kombination av mekanisk och enzymatisk behandling av lungvävnad gav en hög andel lönsamma leukocyter och endotelceller. Närmare bestämt var den mekaniska upplösning av vävnad uppnås genom gentleMACS Dissociator. Den gentleMACS C Rör inkluderar en rotor - stator, som dissocierar vävnad i ett skonsamt sätt. Förfarandet styrs av programmets inställningar av instrumentet. Inställningarna har optimerats för att u...

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		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
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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>
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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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<tr>
<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">Beredningen av encelliga suspensioner från vävnader är en viktig förutsättning för många experiment i cellulära forskning. Processen att det skiljer hela organ kräver specifika parametrar för att få ett stort antal livskraftiga celler på ett reproducerbart sätt. Den gentleMACS Dissociator optimerar denna uppgift med en enkel, praktisk protokoll. Instrumentet innehåller förprogrammerade inställningar som är optimerade för effektiv men skonsam dissociation av olika vävnadstyper, inklusive mus lungorna. I denna publikation användningen av gentleMACS Dissociator på lungvävnad från möss kan påvisas.</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">Ta avstånd celler från specifika vävnadstyper kräver särskild parametrar för vävnad agitation för att få en stor mängd livskraftiga, odlingsbara celler. Den Miltenyi gentleMACS dissociator optimerar denna uppgift med en enkel, praktisk protokoll. I denna publikation användning av denna apparat på lungvävnad förklaras.</p>

Standardiserade Beredning av encelliga Suspensioner från mus lungvävnad med gentleMACS Dissociator

The preparation of single-cell suspensions from tissues is an important prerequisite for many experiments in cellular research. The process of ...

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Carotid arteries are major blood vessels in the neck that supply blood and oxygen to the brain, but carotid stenosis occurs when carotid arteries are ...