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"> Fondsen uit Congressionally Geregisseerd Medical Research Fund-subsidie ​​(DAMD17-01-0264) naar ML Cutler en de Verenigde Staten militaire Kanker Instituut naar ML Cutler wordt ondersteund het werk. </p>

<ol><li> De HC11 cellen werden gekweekt tot confluentie gedurende 6 dagen in RPMI 1640-medium aangevuld met 10% foetaal kalf serum, 5μg/ml insuline, 10 mM HEPES, 10ng/ml epidermale groeifactor (EGF) de bekwaamheid vast te stellen. </li><li> De cellen werden gewassen met PBS en onderhouden groei media zonder EGF voor 24 uur. </li><li> Voor het induceren van lactogenic differentiatie van de cellen werden geïncubeerd in differentiatie media, dat wil zeggen RPMI met dexamethason (10 <sup>-6</sup> M), insuline (5 ug / ml) en prolactine (5 ug / ml)...

<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"> Om deze techniek succesvol te zijn en van de beste kwaliteit, de epitheliale cellen moet worden gehandhaafd op samenloop alvorens te worden geïnduceerd met lactogenic hormonen. Er kunnen situaties zijn waarin er talrijke mammospheres, waardoor de kwantificering moeilijk. Dus, voor kwantitatieve doeleinden is het best om de mammospheres fotograferen op een tijdstip als het aantal mammospheres is in staat moet worden geteld. </p><p class="jove_content"> Hoewel er verschillende moleculaire merkers worden gebruikt om de ma...

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mouse mammary epithelial cells form mammospheres during lactogenic differentiation

<p class="jove_content">Een fenotypische die vaak wordt gebruikt om de mate van lactogenic differentiatie in muis borstklieren epitheelcellen culturen te bepalen, is de vorming van koepelvormige celstructuren aangeduid als mammospheres<sup&gt; 1</sup&gt;. De HC11 cellijn is werkzaam als een modelsysteem voor de studie van de regulering van mammaire lactogenic differentiatie zowel<em&gt; In vitro</em&gt; En<em&gt; In vivo</em<sup&gt; 2</sup&gt;. De HC11 cellen differentiëren en melkeiwitten synthetiseren in reactie op de behandeling met lactogenic hormonen. Naar aanleiding van de groei van de HC11 muis mamma-epitheliale cellen tot confluentie, was lactogenic differentiatie geïnduceerd door de toevoeging van een combinatie van lactogenic hormonen waaronder dexamethason, insuline, en prolactine, aangeduid als DIP. De HC11 cellen geïnduceerd om te differentiëren werden gefotografeerd soms tot 120 uur na inductie van differentiatie en het aantal mammospheres dat verscheen in elke cultuur werd opgesomd. De grootte van de individuele mammospheres correleert met de mate van differentiatie en dit is afgebeeld in de beelden van de differentiërende cellen.</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 differentiatie kan worden gekenmerkt door de vorming van overkoepelde bedoelde structuren als mammospheres. De structuren kunnen worden opgesomd door de fase-contrast-microscopie om te helpen bij het kwantificeren van lactogenic differentiatie.</p>

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