Name: mouse mammary epithelial cells form mammospheres during lactogenic differentiation
Uploaded: 2009-10-06T16:30:00
Description: Watch this Scientific Journal Video about Mouse Mammary Epithelial Cells form Mammospheres During Lactogenic Differentiation at JoVE.com

<p class="jove_content"> Medel från congressionally Regi Medical Research Fund bidrag (DAMD17-01-0264) till ML Cutler och USA militära Cancer Institute till ML Cutler stöttat arbetet. </p>

<ol><li> Den HC11 cellerna odlades till sammanflödet i 6 dagar i RPMI 1640 mediet kompletteras med 10% fetalt kalvserum, 5μg/ml Insulin, 10 mm HEPES, 10ng/ml epidermal tillväxtfaktor (EGF) för att upprätta kompetens. </li><li> Cellerna tvättas med PBS och underhållas i tillväxt media utan fonden i 24 timmar. </li><li> Att förmå lactogenic differentiering cellerna inkuberades i differentiering media, dvs RPMI med dexametason (10 <sup>-6</sup> M), insulin (5 mikrogram / ​​ml) och prolaktin (5 mikrogram / ​​ml) kallas DIP. </li><...

<ol>
	<li>Blatchford, D. R., Hendry, K. A., Turner, M. D., Burgoyne, R. D., Wilde, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vectorial+secretion+by+constitutive+and+regulated+secretory+pathways+in+mammary+epithelial+cells.">Vectorial secretion by constitutive and regulated secretory pathways in mammary epithelial cells.</a> <em style='font-style: italic'>Epithelial Cell Biol</em>. <b>4</b>, 8-16 (1995).</li><li>Wang, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glucocorticoid+induced+expression+of+connective+tissue+growth+factor+contributes+to+lactogenic+differentiation+of+mouse+mammary+epithelial+cells.">Glucocorticoid induced expression of connective tissue growth factor contributes to lactogenic differentiation of mouse mammary epithelial cells.</a> <em style='font-style: italic'>J Cell Physiol</em>. <b>214</b>, 38-46 (2008).</li></ol>

<p class="jove_content"> För att denna teknik för att vara framgångsrika och av bästa kvalitet, måste epitelceller hållas på sammanflödet innan de framkallas med lactogenic hormoner. Det kan finnas situationer där det finns många mammospheres, vilket gör kvantifieringen svår. Således, för kvantitativ ändamål det är bäst att fotografera mammospheres vid en tidpunkt då antalet mammospheres kan räknas. </p><p class="jove_content"> Även om det finns olika molekylära markörer används för att bestämma graden av lactogenic di...

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mouse mammary epithelial cells form mammospheres during lactogenic differentiation

<p class="jove_content">En fenotypiska mått används ofta för att bestämma graden av lactogenic differentiering i mus bröst epitelceller kulturer är bildandet av kupolformade cellstrukturer kallas mammospheres<sup&gt; 1</sup&gt;. Den HC11 cellinje har varit anställd som ett modellsystem för att studera regleringen av bröst lactogenic differentiering både<em&gt; In vitro</em&gt; Och<em&gt; In vivo</em<sup&gt; 2</sup&gt;. Den HC11 cellerna differentiera och syntetisera mjölkprotein som svar på behandling med lactogenic hormoner. Efter ökningen av HC11 mus bröst epitelceller till sammanflödet var lactogenic differentiering framkallade genom tillsats av en kombination av lactogenic hormoner inklusive dexametason, insulin och prolaktin, kallad DIP. Den HC11 celler förmås att skilja fotograferades tidvis upp till 120 timmar efter induktion av differentiering och antalet mammospheres som dök upp i varje kultur räknades. Storleken på de enskilda mammospheres korrelerar med graden av differentiering och detta skildras i bilderna av de differentierande celler.</p>

Watch this Scientific Journal Video about Mouse Mammary Epithelial Cells form Mammospheres During Lactogenic Differentiation at JoVE.com

Mouse Mammary Epithelial Cells form Mammospheres During Lactogenic Differentiation

<p class="jove_content">HC11 lactogenic differentiering kan karakteriseras genom bildandet av välvda strukturer som kallas mammospheres. Strukturerna kan räknas upp faskontrast mikroskopi till stöd i kvantifiera lactogenic differentiering.</p>

Mus bröst epitelceller formen Mammospheres Under Lactogenic Differentiering

A phenotypic measure commonly used to determine the degree of lactogenic differentiation in mouse mammary epithelial cell cultures is the formation of ...

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