Name: isolation of circulating tumor cells in an orthotopic mouse model of colorectal cancer
Uploaded: 2017-07-18T13:00:00
Description: Watch this Scientific Journal Video about Isolation of Circulating Tumor Cells in an Orthotopic Mouse Model of Colorectal Cancer at JoVE.com

Ce travail a été soutenu par la fondation allemande de recherche (WE 3548 / 4-1) et Roland-Ernst-Stiftung für Gesundheitswesen (1/14).

Les expériences sur les animaux présentées ici ont été revues et autorisées indépendamment par un comité institutionnel et gouvernemental de soins et d&#39;utilisation des animaux et ont été menées selon les directives de la Fédération des associations de scientifiques de laboratoire (FELASA). Toutes les mesures possibles ont été prises pour minimiser les souffrances, y compris l&#39;anesthésie et l&#39;analgésie ou, si nécessaire, une euthanasie prématurée. 1. Pr?...

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N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Meta-analysis+shows+that+detection+of+circulating+tumor+cells+indicates+poor+prognosis+in+patients+with+colorectal+cancer.">Meta-analysis shows that detection of circulating tumor cells indicates poor prognosis in patients with colorectal cancer.</a> Gastroenterology. 138 (5), 1714-1726 (2010).</li><li>Steinert, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immune+Escape+and+Survival+Mechanisms+in+Circulating+Tumor+Cells+of+Colorectal+Cancer.">Immune Escape and Survival Mechanisms in Circulating Tumor Cells of Colorectal Cancer.</a> Cancer Res. 74 (6), 1694-1704 (2014).</li><li>Sharpless, N. E., Depinho, R. 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E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=TLR+activation+and+ionizing+radiation+induce+strong+immune+responses+against+multiple+tumor+entities.">TLR activation and ionizing radiation induce strong immune responses against multiple tumor entities.</a> Oncoimmunology. 4 (11), e1042201 (2015).</li><li>Sch&#246;lch, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Circulating+tumor+cells+exhibit+stem+cell+characteristics+in+an+orthotopic+mouse+model+of+colorectal+cancer.">Circulating tumor cells exhibit stem cell characteristics in an orthotopic mouse model of colorectal cancer.</a> Oncotarget. 7 (19), 27232-27242 (2016).</li><li>Nanduri, L. 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Malgré leur activité précliniquement prouvée dans les modèles de souris sous-cutanés, la grande majorité des nouveaux composés échouent dans les essais cliniques et n&#39;atteignent jamais la clinique. 11 Cette insuffisance évidente de modèles de souris sous-cutanés pour simuler de manière précise la biologie et les modèles de croissance des tumeurs a conduit au développement de modèles de souris orthotopiques basés sur l&#39;injection de cellules tumorales directement dans l&#3...

Le cancer colorectal (CRC) est l&#39;une des causes les plus fréquentes de décès par cancer dans les pays occidentaux. 1 Alors que la tumeur primaire peut souvent être réséquée, l&#39;apparition de métastases à distance aggrave considérablement le pronostic et conduit souvent à la mort. 2 , 3 La corrélation biologique de la métastase est la circulation des cellules tumorales (CTC), qui se détachent de la tumeur, survivent en circulation, attachent à l&#39;épithélium dans l&#39;organe cible, envahissent l&#39;organe et finissent par dépasser de nouvelles lésions. 4 Bien que les CTC connaissent une pertinence pronostique, 5 , 6 , 7 , 8 , 9, leur biologie n&#39;est que partiellement comprise en raison de leur extrême rareté dans le CRC. dix Les modèles de souris sont un puissant tOol pour étudier divers aspects de la biologie du cancer. Les modèles classiques de tumeur sous-cutanée sont produits par injection sous-cutanée de cellules tumorales dans des souris destinataires, qui peuvent être immunocompétentes (si des cellules tumorales murines syngéniques sont utilisées) ou immunodéficientes. Les modèles de tumeur sous-cutanée sont peu coûteux et produisent rapidement des données; Leur croissance tumorale de point final peut être mesurée de manière simple et non invasive. Cependant, 88% des nouveaux composés qui ont démontré une activité antitumorale dans de tels modèles échouent dans les essais cliniques. 11 Cela s&#39;explique en partie par les différences interspécifiques entre les humains et les souris; Cependant, une grande partie de cette défaillance est due à la faible valeur prédictive des modèles de souris sous-cutanés. Les modèles de souris orthotopiques, dans lesquels les cellules tumorales sont injectées dans l&#39;organe d&#39;origine et donc poussent dans leur microenvironnement original, sont de plus en plus utilisés dans la recherche sur le cancer. 11 , 12 , </sUp&gt; 13 , 14 Les modèles orthotopiques ne simulent pas seulement les conditions locales de croissance tumorale; En raison du site anatomiquement correct de la croissance de la tumeur, les modèles de souris orthotopiques permettent également une simulation réaliste des métastases et sont donc utilisés pour étudier la biologie CTC 8 , 15 , 16 ou leur réponse à différents traitements dans le CRC. 13 , 17 Un inconvénient majeur des modèles de souris orthotopiques est leur complexité technique. Selon l&#39;organe dans lequel les cellules doivent être injectées, la courbe d&#39;apprentissage jusqu&#39;à ce que l&#39;expérimentateur puisse induire des tumeurs reproductibles soit assez longue. Cela s&#39;applique particulièrement aux modèles de cancer colorectal, car les cellules tumorales doivent être injectées dans la paroi intestinale, ce qui entraîne souvent une perforation, une fuite de cellules tumorales ou une perte de cellules tumorales endoluminales. C&#39;est unL&#39;article a pour but de décrire le procédé de préparation cellulaire à partir d&#39;échantillons de tissus primaires, de lignées cellulaires 2D et de culture organo-3D et leur injection dans le cécum des souris. La technique décrite ici conduit à des tumeurs hautement uniformes et, en fonction de la biologie tumorale de la lignée cellulaire utilisée pour l&#39;injection, de la formation reproductible de métastases à distance et de CTC chez la souris destinataire. 15

<table border="0" cellpadding="0" cellspacing="0" width="1294">
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		<tr height="25">
			<td height="25" style="height:25px;width:421px;">Cell culture Media and Components</td>
			<td style="width:297px;"></td>
			<td style="width:119px;"></td>
			<td style="width:457px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Advanced DMEM F12</td>
			<td style="width:297px;">Invitrogen</td>
			<td style="width:119px;">12634010</td>
			<td style="width:457px;">DMEM/ F12 +++ medium</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">HEPES (1 M)</td>
			<td style="width:297px;">Life Technologies GmbH</td>
			<td style="width:119px;">15630056</td>
			<td style="width:457px;">DMEM/ F12 +++ medium</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Glutamax-I Supplement (200 mM)</td>
			<td style="width:297px;">Life Technologies GmbH</td>
			<td style="width:119px;">35050038</td>
			<td style="width:457px;">DMEM/ F12 +++ medium</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Penicillin/Streptomycin (PenStrep)</td>
			<td style="width:297px;">Life Technologies GmbH</td>
			<td style="width:119px;">15140122</td>
			<td style="width:457px;">DMEM/ F12 +++ medium</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">DMEM</td>
			<td style="width:297px;">Life Technologies GmbH</td>
			<td style="width:119px;">61965026</td>
			<td>basic medium of 2D cell lines (DMEM/10%FCS)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Fetal Calf Serum (FCS)</td>
			<td style="width:297px;">BIOCHROM AG</td>
			<td style="width:119px;">S 0115</td>
			<td>basic medium of 2D cell lines (DMEM/10%FCS)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">TrypLE Express enzymatic dissociation buffer</td>
			<td style="width:297px;">Life Technologies GmbH</td>
			<td style="width:119px;">12604021</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Matrigel basement membrane matrix (BMM, phenol red free)</td>
			<td style="width:297px;">CORNING B.V. Life Sciences</td>
			<td style="width:119px;">356231</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Dulbecco&#39;s Phosphate Buffered Saline</td>
			<td style="width:297px;">Life Technologies GmbH</td>
			<td style="width:119px;">14190169</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Trypsin-EDTA (0,25%, Phenol-Red)</td>
			<td style="width:297px;">Life Technologies GmbH</td>
			<td style="width:119px;">25200072</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">6-/48-well plates with lid</td>
			<td style="width:297px;">CORNING</td>
			<td style="width:119px;">3516/3548</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">cell culture flask 75cm&#178;, 250 mL</td>
			<td style="width:297px;">VWR International GmbH</td>
			<td style="width:119px;">734-2066</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">cell culture flask 150cm&#178;, 600 mL</td>
			<td style="width:297px;">Corning B.V. Life Sciences</td>
			<td style="width:119px;">355001</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:421px;">Eppendorf tubes 1,5 mL / 2 mL</td>
			<td style="width:297px;">Sarstedt AG &#38; Co.</td>
			<td style="width:119px;">72.706.400/ 72.695.400</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">15 ml, 50 ml centrifuge tubes</td>
			<td style="width:297px;">Greiner-Bio-One GmbH</td>
			<td style="width:119px;">188271/227270</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">TC10 Counting Slides (for TC20 Counting Machine)</td>
			<td style="width:297px;">Bio-Rad Laboratories GmbH</td>
			<td style="width:119px;">1450016</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:421px;">Pasteur pipettes (glass, 150 mm)</td>
			<td style="width:297px;">Fisher Scientific GmbH</td>
			<td style="width:119px;">11546963/ FB50251</td>
			<td>thinly pulled by using a bunsen burner</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">gentleMACS Dissociator</td>
			<td style="width:297px;">Miltenyi Biotec</td>
			<td>130-093-235</td>
			<td>for primary tumor tissue preparation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">MACSmix Tube Rotator</td>
			<td style="width:297px;">Miltenyi Biotec</td>
			<td>130-090-753</td>
			<td>for primary tumor tissue preparation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">gentleMACS C Tubes</td>
			<td style="width:297px;">Miltenyi Biotec</td>
			<td>130-093-237</td>
			<td>for primary tumor tissue preparation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Human Tumor Dissociation Kit</td>
			<td style="width:297px;">Miltenyi Biotec</td>
			<td>130-095-929</td>
			<td>for primary tumor tissue preparation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Falcon 70&#181;m Cell Strainer</td>
			<td style="width:297px;">Corning B.V. Life Sciences</td>
			<td>352350</td>
			<td>for primary tumor tissue preparation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Name</td>
			<td style="width:297px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:421px;">Surgical Equipment</td>
			<td style="width:297px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Sevoflurane</td>
			<td style="width:297px;">AbbVie Germany GmbH &#38; Co. KG</td>
			<td style="width:119px;">-</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Medical oxygen</td>
			<td style="width:297px;">Air Liquide Medical GmbH</td>
			<td style="width:119px;">-</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Buprenorphine</td>
			<td style="width:297px;">Temgesic</td>
			<td style="width:119px;">-</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Bepanthen - opthalmic ointment</td>
			<td style="width:297px;">Bayer Vital GmbH</td>
			<td style="width:119px;">10047757</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Normal saline 0.9% (E154)</td>
			<td style="width:297px;">Serumwerk Bernburg AG</td>
			<td style="width:119px;">10013</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Aqua ad injectabilia</td>
			<td style="width:297px;">Braun</td>
			<td style="width:119px;">235144</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">1 mL Syringe (without dead volume) - Injekt-F SOLO</td>
			<td style="width:297px;">Braun/neoLab</td>
			<td style="width:119px;">194291661</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">30G injection needle</td>
			<td style="width:297px;">BECTON DICKINSON</td>
			<td style="width:119px;">304000</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">cellulose swabs</td>
			<td style="width:297px;">Lohmann &#38; Rauscher Deutschland</td>
			<td style="width:119px;">13356</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Micro-Adson Forceps</td>
			<td style="width:297px;">FST - Fine Science Tools</td>
			<td style="width:119px;">11018-12</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Iris Scissor - ToughCut</td>
			<td style="width:297px;">FST - Fine Science Tools</td>
			<td style="width:119px;">14058-11</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Olsen-Hegar Needle Holder</td>
			<td style="width:297px;">FST - Fine Science Tools</td>
			<td style="width:119px;">12002-12</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">AutoClip Kit</td>
			<td style="width:297px;">FST - Fine Science Tools</td>
			<td style="width:119px;">12020-00</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">PDS Z1012H 6/0 C1 (surgical suture)</td>
			<td style="width:297px;">Johnson &#38; Johnson Medical GmbH</td>
			<td style="width:119px;">Z1012H</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:421px;">Table Top Research Anesthesia Machine w/O2 Flush and a Sevoflurane Vaporizer</td>
			<td style="width:297px;">Parkland Scientific</td>
			<td style="width:119px;">V3000PS/PK</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">UltraMicro Pump with Micro4 Controller</td>
			<td style="width:297px;">World Precision Instruments</td>
			<td style="width:119px;">UMP3-4</td>
			<td>equipment for highly controlled orthotopic injection</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Footswitch for SYS-Micro4 Controller</td>
			<td style="width:297px;">World Precision Instruments</td>
			<td style="width:119px;">15867</td>
			<td>equipment for highly controlled orthotopic injection</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Three-axis Manual Micromanipulator</td>
			<td style="width:297px;">World Precision Instruments</td>
			<td style="width:119px;">M325</td>
			<td>equipment for highly controlled orthotopic injection</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Magnetic Stand for Micromanipulator</td>
			<td style="width:297px;">World Precision Instruments</td>
			<td style="width:119px;">M10</td>
			<td>equipment for highly controlled orthotopic injection</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Steel Base Plate for M10 Magnetic Stand</td>
			<td style="width:297px;">World Precision Instruments</td>
			<td style="width:119px;">5479</td>
			<td>equipment for highly controlled orthotopic injection</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Hot Plate 062</td>
			<td style="width:297px;">Labotect</td>
			<td style="width:119px;">13854</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Isis - Hair shaver</td>
			<td style="width:297px;">AESCULAP - Braun</td>
			<td style="width:119px;">-</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Binocular Surgical Microscope</td>
			<td style="width:297px;">Parkland Scientific</td>
			<td style="width:119px;">VS-2Z</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Name</td>
			<td style="width:297px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:421px;">CTC isolation</td>
			<td style="width:297px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">EDTA</td>
			<td style="width:297px;">Roth</td>
			<td>8040.1</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Density gradient medium &#8211; Ficoll</td>
			<td>StemCell - Lymphoprep</td>
			<td>7801</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:421px;">Alexa Fluor 488 anti-human CD326 (EpCAM) Antibody clone 9C4</td>
			<td>BioLegend</td>
			<td>324210</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:421px;">Alexa Fluor 488 anti-mouse CD326 (EpCAM) Antibody clone G8.8</td>
			<td>BioLegend</td>
			<td style="width:119px;">118210</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Petri Dish, &#248; 60 x 15 mm, 21 cm&#178;, Vent</td>
			<td>Greiner bio-one</td>
			<td>628102</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fluorescence Cell Culture Microscope</td>
			<td style="width:297px;">Leica</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Transferman 4r Micromanipulator</td>
			<td style="width:297px;">Eppendorf</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">CellTram Air</td>
			<td style="width:297px;">Eppendorf</td>
			<td style="width:119px;"></td>
			<td style="width:457px;">aspiration pump connected to the micromanipulator</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dmz Universal Microelectrode Puller</td>
			<td style="width:297px;">Dagan Corporation</td>
			<td style="width:119px;"></td>
			<td>required for the manufacturing of micro capillaries for single cell aspiration</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Prism Glass Capillaries</td>
			<td style="width:297px;">Dagan Corporation</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">PAP pen</td>
			<td style="width:297px;">Abcam</td>
			<td style="width:119px;">ab2601</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Dulbecco&#39;s Phosphate Buffered Saline</td>
			<td style="width:297px;">Life Technologies GmbH</td>
			<td style="width:119px;">14190169</td>
			<td>picking buffer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Fetal Calf Serum (FCS)</td>
			<td style="width:297px;">BIOCHROM AG</td>
			<td style="width:119px;">S 0115</td>
			<td>picking buffer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Penicillin/Streptomycin (PenStrep)</td>
			<td style="width:297px;">Life Technologies GmbH</td>
			<td style="width:119px;">15140122</td>
			<td>picking buffer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">EDTA</td>
			<td style="width:297px;">Roth</td>
			<td>8040.1</td>
			<td>picking buffer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Name</td>
			<td style="width:297px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:421px;">Immunohistochemistry</td>
			<td style="width:297px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">Purified anti-human CD326 (EpCAM) antibody clone 9C4</td>
			<td>BioLegend</td>
			<td style="width:119px;">324201</td>
			<td>EpCAM immunohistochemistry (cf, fig 2C)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:421px;">HRP rabbit anti-mouse IgG</td>
			<td style="width:297px;">Abcam</td>
			<td style="width:119px;">ab97046</td>
			<td>EpCAM immunohistochemistry (cf, fig 2C)</td>
		</tr>
	</tbody>
</table>

isolation of circulating tumor cells in an orthotopic mouse model of colorectal cancer

Malgré les avantages d&#39;une applicabilité et d&#39;une rentabilité faciles, les modèles de souris sous-cutanés ont de graves limites et ne simulent pas exactement la biologie tumorale et la dissémination de cellules tumorales. Des modèles de souris orthotopiques ont été introduits pour surmonter ces limites; Cependant, ces modèles sont techniquement exigeants, en particulier dans les organes creux tels que le gros intestin. Afin de produire des tumeurs uniformes qui se développent de manière fiable et de métastases, des techniques normalisées de préparation et d&#39;injection de cellules tumorales sont essentielles. Nous avons développé un modèle orthotopique de cancer colorectal (CRC) qui développe des tumeurs hautement uniformes et peut être utilisé pour des études de biologie tumorale ainsi que des essais thérapeutiques. Les cellules tumorales provenant des tumeurs primaires, des lignées cellulaires bidimensionnelles (2D) ou des organoïdes tridimensionnels (3D) sont injectées dans le cécum et, selon le potentiel métastatique des cellules tumorales injectées, forment des tumeurs hautement métastatiques. En outre,Les CTC peuvent être consultés régulièrement. Nous décrivons ici la technique de la préparation des cellules tumorales à partir des lignées cellulaires 2D et des organoïdes 3D, ainsi que du tissu tumoral primaire, des techniques chirurgicales et d&#39;injection ainsi que de l&#39;isolement des CTC des souris portant une tumeur et des conseils pour le dépannage.

La génération réussie et reproductible de tumeurs colorectales dans ce modèle dépend de manière critique d&#39;une injection précise des cellules sans déversement ou fuite. Si cela est atteint, ce modèle est extrêmement fiable et entraîne très rarement une diffusion péritonéale artificielle. La cinétique de croissance des tumeurs ainsi que leurs profils de diffusion dépendent de la biologie des organoïdes et des cellules utilisés. 15 Alors que le...

Watch this Scientific Journal Video about Isolation of Circulating Tumor Cells in an Orthotopic Mouse Model of Colorectal Cancer at JoVE.com

Isolation of Circulating Tumor Cells in an Orthotopic Mouse Model of Colorectal Cancer

Nous décrivons l&#39;établissement de tumeurs colorectales orthotopiques par injection de cellules tumorales ou organoïdes dans le cécum des souris et l&#39;isolement subséquent des cellules tumorales circulantes (CTC) de ce modèle.

Isolation des cellules tumorales circulantes dans un modèle de souris orthotopique du cancer colorectal

Despite the advantages of easy applicability and cost-effectiveness, subcutaneous mouse models have severe limitations and do not accurately simulate ...

Research

JoVE Journal

Cancer Research

Fluorescent Orthotopic Mouse Model of Pancreatic Cancer

Fluorescent modèle de souris orthotopique de cancer du pancréas

Pancreatic cancer remains one of the cancers for which survival has not improved substantially in the last few decades. Only 7% of diagnosed patients will ...

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Recherche en cancérologie

Murine Experimental Model of Original Tumor Development and Peritoneal Metastasis via Orthotopic Inoculation with Ovarian Carcinoma Cells

Murine Experimental Model of Original Tumor Development and Peritoneal Metastasis via Orthotopic Inoculation with Ovarian Carcinoma Cells

Murin modèle expérimental de tumeur de développement original et péritonéale Métastase<em&gt; via</em&gt; Orthotopique Inoculation avec des cellules de l&#39;ovaire Carcinome

Epithelial ovarian carcinoma (EOC) is associated with a poor prognosis because it shows peritoneal dissemination. To improve the prognosis, it is ...

Capture and Release of Viable Circulating Tumor Cells from Blood

Captage et sortie de Viable cellules tumorales circulantes du sang

We demonstrate a method for size based capture of viable circulating tumor cell (CTC) from whole blood, along with the release of these cells from chip ...

A Murine Orthotopic Bladder Tumor Model and Tumor Detection System

Un système de détection de tumeur modèle et tumeur murine Orthotopique vessie

This protocol describes the generation of bladder tumors in female C57BL/6J mice using the murine bladder cancer cell line MB49, which has been modified ...

A Genetically Engineered Mouse Model of Sporadic Colorectal Cancer

Un modèle de souris génétiquement modifiée de cancer colorectal sporadique

Despite the advantages of easy applicability and cost-effectiveness, colorectal cancer mouse models based on tumor cell injection have severe limitations ...

Isolation of Human Endothelial Cells from Normal Colon and Colorectal Carcinoma - An Improved Protocol

Isolement des cellules endothéliales humaines du côlon Normal et carcinome Colorectal - un protocole amélioré

Primary cells isolated from human carcinomas are valuable tools to identify pathogenic mechanisms contributing to disease development and progression. In ...

Detection of a Circulating MicroRNA Custom Panel in Patients with Metastatic Colorectal Cancer

Détection d’un Panel de micro-ARN Custom circulant chez les Patients atteints de Cancer Colorectal métastatique

There is increasing interest in liquid biopsy for cancer diagnosis, prognosis and therapeutic monitoring, creating the need for reliable and useful ...

Micromanipulation of Circulating Tumor Cells for Downstream Molecular Analysis and Metastatic Potential Assessment

Micromanipulation de cellules tumorales circulantes pour l’analyse moléculaire en aval et l’évaluation du potentiel métastatique

Blood-borne metastasis accounts for&#160;most cancer-related deaths and involves circulating tumor cells (CTCs) that are successful in establishing new ...

Semi-automatic PD-L1 Characterization and Enumeration of Circulating Tumor Cells from Non-small Cell Lung Cancer Patients by Immunofluorescence

Caractérisation semi-automatique De MD-L1 et recensement des cellules tumorales circulantes de patients atteints d'immunofluorescence

Circulating tumor cells (CTCs) derived from the primary tumor are shed into the bloodstream or lymphatic system. These rare cells (1&#8722;10 cells per mL ...

Culture, Manipulation, and Orthotopic Transplantation of Mouse Bladder Tumor Organoids

Culture, Manipulation et Transplantation orthotopique d'organoïdes tumoraux de la vessie de souris

The development of advanced tumor models has long been encouraged because current cancer models have shown limitations such as lack of three-dimensional ...