Name: generation of high-throughput three-dimensional tumor spheroids for drug screening
Uploaded: 2018-09-05T15:30:00
Description: Watch this Scientific Journal Video about Generation of High-Throughput Three-Dimensional Tumor Spheroids for Drug Screening at JoVE.com

Forfatterne vil gerne takke Marc Ferrer på National Center for fremme translationel videnskab, National Institutes of Health for hans støtte og vejledning til den indledende udvikling af disse assays. Derudover vil vi gerne takke Alyson Freeman, Mariela Jaskelioff, Michael Acker, Jacob Haling og Vesselina Cooke for deres videnskabelige input og drøftelser som projekt teammedlemmer.

1. dyrkning af 1536-godt 3D Tumor Spheroids
<ol><li>Forberede en frisk 3D tumor klumpformet medium stilk, (celle medium + knockout serum udskiftning + insulin-transferrin-selen) ved at tilføje de følgende reagenser til 500 mL af Dulbeccos modificerede Eagles Medium (DMEM/F12): 1 x penicillin/streptomycin, 10 ng/mL af human Basic fibroblast vækstfaktor (bFGF), 20 ng/mL af human epidermal vækstfaktor (EGF), 0,4% bovint serumalbumin (BSA) (brøkdel V), 1 x insulin-transferrin-selen, og 1% knockout (KO) serum udskiftni...

<ol>
	<li>Lee, M., Vasioukhin, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+polarity+and+cancer--cell+and+tissue+polarity+as+a+non-canonical+tumor+suppressor.">Cell polarity and cancer--cell and tissue polarity as a non-canonical tumor suppressor.</a> Journal of Cell Science. 121 (Pt 8), 1141-1150 (2008).</li><li>Nath, S., Devi, G. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-dimensional+culture+systems+in+cancer+research:+Focus+on+tumor+spheroid+model.">Three-dimensional culture systems in cancer research: Focus on tumor spheroid model.</a> Pharmacology &amp; Therapeutics. 163, 94-108 (2016).</li><li>Weigelt, B., Ghajar, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+need+for+complex+3D+culture+models+to+unravel+novel+pathways+and+identify+accurate+biomarkers+in+breast+cancer.">The need for complex 3D culture models to unravel novel pathways and identify accurate biomarkers in breast cancer.</a> Advanced Drug Delivery Reviews. 69, 42-51 (2014).</li><li>Li, Q., Chow, A. B., Mattingly, R. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-dimensional+overlay+culture+models+of+human+breast+cancer+reveal+a+critical+sensitivity+to+mitogen-activated+protein+kinase+kinase+inhibitors.">Three-dimensional overlay culture models of human breast cancer reveal a critical sensitivity to mitogen-activated protein kinase kinase inhibitors.</a> The Journal of Pharmacology and Experimental Therapeutics. 332 (3), 821-828 (2010).</li><li>Li, L., Lu, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimizing+a+3D+Culture+System+to+Study+the+Interaction+between+Epithelial+Breast+Cancer+and+Its+Surrounding+Fibroblasts.">Optimizing a 3D Culture System to Study the Interaction between Epithelial Breast Cancer and Its Surrounding Fibroblasts.</a> Journal of Cancer. 2, 458-466 (2011).</li><li>Pickl, M., Ries, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+3D+and+2D+tumor+models+reveals+enhanced+HER2+activation+in+3D+associated+with+an+increased+response+to+trastuzumab.">Comparison of 3D and 2D tumor models reveals enhanced HER2 activation in 3D associated with an increased response to trastuzumab.</a> Oncogene. 28 (3), 461-468 (2009).</li><li>Nyga, A., Cheema, U., Loizidou, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=3D+tumour+models:+novel+in+vitro+approaches+to+cancer+studies.">3D tumour models: novel in vitro approaches to cancer studies.</a> Journal of Cell Communication and Signaling. 5 (3), 239-248 (2011).</li><li>Kimlin, L. C., Casagrande, G., Virador, V. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vitro+three-dimensional+(3D)+models+in+cancer+research:+an+update.">In vitro three-dimensional (3D) models in cancer research: an update.</a> Molecular Carcinogenesis. 52 (3), 167-182 (2013).</li><li>Mathews Griner, L. A., 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=Large-scale+pharmacological+profiling+of+3D+tumor+models+of+cancer+cells.">Large-scale pharmacological profiling of 3D tumor models of cancer cells.</a> Cell Death &amp; Disease. 7 (12), e2492 (2016).</li><li>Polo, M. L., 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=Responsiveness+to+PI3K+and+MEK+inhibitors+in+breast+cancer.+Use+of+a+3D+culture+system+to+study+pathways+related+to+hormone+independence+in+mice.">Responsiveness to PI3K and MEK inhibitors in breast cancer. Use of a 3D culture system to study pathways related to hormone independence in mice.</a> PLoS One. 5 (5), e10786 (2010).</li><li>Hatzivassiliou, 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=RAF+inhibitors+prime+wild-type+RAF+to+activate+the+MAPK+pathway+and+enhance+growth.">RAF inhibitors prime wild-type RAF to activate the MAPK pathway and enhance growth.</a> Nature. 464 (7287), 431-435 (2010).</li><li>Bhargava, A., 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=Registered+report:+RAF+inhibitors+prime+wild-type+RAF+to+activate+the+MAPK+pathway+and+enhance+growth.">Registered report: RAF inhibitors prime wild-type RAF to activate the MAPK pathway and enhance growth.</a> Elife. 5, e09976 (2016).</li><li>Mathews, L. A., Hurt, E. M., Zhang, X., Farrar, W. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epigenetic+regulation+of+CpG+promoter+methylation+in+invasive+prostate+cancer+cells.">Epigenetic regulation of CpG promoter methylation in invasive prostate cancer cells.</a> Molecular Cancer. 9, 267 (2010).</li><li>Mathews, L. A., Crea, F., Farrar, W. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epigenetic+gene+regulation+in+stem+cells+and+correlation+to+cancer.">Epigenetic gene regulation in stem cells and correlation to cancer.</a> Differentiation. 78 (1), 1-17 (2009).</li><li>Mathews, L. A., 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=Increased+expression+of+DNA+repair+genes+in+invasive+human+pancreatic+cancer+cells.">Increased expression of DNA repair genes in invasive human pancreatic cancer cells.</a> Pancreas. 40 (5), 730-739 (2011).</li><li>Mathews, L. A., 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=A+1536-well+quantitative+high-throughput+screen+to+identify+compounds+targeting+cancer+stem+cells.">A 1536-well quantitative high-throughput screen to identify compounds targeting cancer stem cells.</a> Journal of Biomolecular Screening. 17 (9), 1231-1242 (2012).</li><li>Horii, T., Nagao, Y., Tokunaga, T., Imai, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Serum-free+culture+of+murine+primordial+germ+cells+and+embryonic+germ+cells.">Serum-free culture of murine primordial germ cells and embryonic germ cells.</a> Theriogenology. 59 (5-6), 1257-1264 (2003).</li><li>Sun, L., 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=Epigenetic+regulation+of+SOX9+by+the+NF-kappaB+signaling+pathway+in+pancreatic+cancer+stem+cells.">Epigenetic regulation of SOX9 by the NF-kappaB signaling pathway in pancreatic cancer stem cells.</a> Stem Cells. 31 (8), 1454-1466 (2013).</li><li>Mathews, L. A., 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=A+1536-well+quantitative+high-throughput+screen+to+identify+compounds+targeting+cancer+stem+cells.">A 1536-well quantitative high-throughput screen to identify compounds targeting cancer stem cells.</a> Journal of Biomolecular Screening. 17 (9), 1231-1242 (2012).</li><li>Mathews Griner, L. A., 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=High-throughput+combinatorial+screening+identifies+drugs+that+cooperate+with+ibrutinib+to+kill+activated+B-cell-like+diffuse+large+B-cell+lymphoma+cells.">High-throughput combinatorial screening identifies drugs that cooperate with ibrutinib to kill activated B-cell-like diffuse large B-cell lymphoma cells.</a> Proceedings of the National Academy of Sciences of the United States of America. 111 (6), 2349-2354 (2014).</li><li>Herter, 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=A+novel+three-dimensional+heterotypic+spheroid+model+for+the+assessment+of+the+activity+of+cancer+immunotherapy+agents.">A novel three-dimensional heterotypic spheroid model for the assessment of the activity of cancer immunotherapy agents.</a> Cancer Immunology, Immunotherapy. 66 (1), 129-140 (2017).</li></ol>

De metoder, der præsenteres her dokumentere en detaljeret protokol om, hvordan man producerer tumor spheroids i 1536-godt plader for store sammensatte screeninger. Disse metoder var oprindeligt tilpasset fra arbejde på National Cancer Institute hvor tumor spheroids blev dyrket i lav-overførselshastighed assays i 6-godt og 96-brønd plader til at stille spørgsmål om genetiske afhængigheder og sammensatte følsomhed13, 14 , 15</su...

I det seneste årti, har flere og flere undersøgelser gennemført brugen af 3D celle kultur modeller til at forstå begreber, som ikke er fuldt sammenfattet af voksende celler i 2D på plastik. Eksempler på disse koncepter omfatter vekslen i normale epitelcelle polaritet1 hvor den rumlige orientering af apikale og basal lag af celler går tabt, samt rollen, den ekstracellulære matrix i reguleringen af overlevelse og celle skæbne. Onkologi forskning, navnlig har brugt 3D modeller til at forstå grundlæggende biologi kræftceller og forskellene mellem 2D og 3D celle kultur systemer3,4. Udviklingen af mere avancerede celle kultur teknikker og deres yderligere tilpasning til multi godt formater har aktiveret søgen efter nye stoffer i 3D-indstillinger. I modsætning til celler dyrkes betingelser 2D, 3D-modeller af tumorer varierer i kompleksitet fra lagdelt mobiltelefonsystemer5 til single-celletype områder af forskellige størrelser, til komplekse multi-cell-type kugler6,7, 8. opdagelsen af nye forbindelser eller biologics, der potent inducerer celledød i disse 3D-modelsystemer er derfor af stor interesse i stof udvikling kampagner. Slutpunkterne på disse assays ofte er identiske med dem som i 2D kulturer til at vurdere ændringer i celledelingen, men da gennemført i en mere fysiologisk relevante indstilling, de kan afsløre den sande niveau af afhængighed af target gen eller pathway bliver afhørt.
Som indført ovenfor, en række modelsystemer er blevet udviklet for at studere medicin svar i 3D kultur systemer, men fleste bruger enten 96 eller 384-godt mikrotiter plader og er ikke let at tilpasse til high throughput screening (HTS) formater ofte bruges i Drug discovery screening kampagner. Sådanne systemer omfatter brugen af hængende droplet teknologier, klumpformet kulturer, pulserende celler med magnetiske partikler til at fremkalde levitation og kulturer indarbejde naturlige eller syntetiske geler som kollagen, Matrigel eller polyethylenglycol (PEG)2 . Her præsenterer vi de detaljerede metoder til en tidligere udviklet teknik til at producere 3D klumpformet kulturer fra etablerede kræft cellelinjer i 1536-godt plade format. I denne protokol, er et meget defineret medium anvendes som forhindrer udlæg i normalt vedhængende celle linjer9. Dette system har begrænsninger (dvs., det kan ikke fuldt sammenfatte en kompleks modelsystem af kræft), men alligevel, disse assays aktiverer en høj overførselshastighed screening af store samlinger af små molekyler og krydsende sammenligninger af narkotika svar mellem 2D og 3D kulturer mod en række cellelinjer og forbindelser.
Cellelinjer valgt for at vise metoder i denne artikel alle harbor mutationer i gener relateret til MAPK signalering vej, en vej, som er meget dysregulated i kræft, og for hvilke mange therapeutics er tilgængelige. Mange af linjerne har aktiveringen muterede mutationer i Kirsten rotte sarkom virus også benævnt KRAS, Neuroblastoma RAS eller de nationale tilsynsmyndigheder, Harvey rotte sarkom virus onkogen eller HRAS og de tilknyttede kinaser hurtigt accelereret fibrosarkomer, også kendt som RAFs. Seneste litteratur tyder på, at forskellige noder af denne vej-hæmmere er entydigt mere virkningsfuldt i et undersæt af cellelinjer, når de dyrkes under 3D betingelser9,10. En undersøgelse viste, at når kræftceller med aktive RAS var kulturperler i 3D, de viste en øget følsomhed over for MAPK hæmmere, og yderligere, at denne tilgang kunne identificere veje og målrettet hæmmere, der kan blive savnet i det traditionelle 2D indstilling. Målet med denne undersøgelse er at præsentere metoder til kultur disse cellelinjer, og yderligere, for at demonstrere differentieret svar til disse hæmmere, som kan observeres, når ved hjælp af 3D celle kultur systemer.

<table>
	<tbody>
		<tr>
			<td>Reagents</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM Dulbecco`s Modified Eagle Media (DMEM)/F12</td>
			<td>ATCC</td>
			<td>30-2006</td>
			<td>Purchased as a prepared solution. This reciepe was found to be preferred compared to other vendors.</td>
		</tr>
		<tr>
			<td>DMEM Dulbecco`s Modified Eagle Media (DMEM)/F12</td>
			<td>Lonza</td>
			<td>12-604F</td>
			<td>Purchased as a prepared solution.</td>
		</tr>
		<tr>
			<td>Roswell Park Memorial Institute (RPMI)</td>
			<td>Lonza</td>
			<td>12-115Q</td>
			<td>Purchased as a prepared solution.</td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum (FBS)</td>
			<td>Seradigm</td>
			<td>1500-500</td>
			<td>Purchased as a prepared solution.</td>
		</tr>
		<tr>
			<td>1x 0.25% Trypsin with Ethylenediaminetetraacetic acid (EDTA)</td>
			<td>Hyclone</td>
			<td>SH30042.01</td>
			<td>Purchased as a prepared solution.</td>
		</tr>
		<tr>
			<td>1x Penicillin/Streptomycin</td>
			<td>Gibco</td>
			<td>15140</td>
			<td>Purchased as a prepared solution.</td>
		</tr>
		<tr>
			<td>10 ng/mL Human basic Fibrobast Growth Factor (bFGF)</td>
			<td>Sigma</td>
			<td>F0291</td>
			<td>Resuspend in sterile water.</td>
		</tr>
		<tr>
			<td>20 ng/mL Human Epidermal Growth Factor (EGF)</td>
			<td>Sigma</td>
			<td>E9644</td>
			<td>Resuspend in sterile water.</td>
		</tr>
		<tr>
			<td>0.4% Bovine Serum Albumin (BSA)</td>
			<td>Sigma</td>
			<td>A9418</td>
			<td>Resuspend in sterile PBS and filter.</td>
		</tr>
		<tr>
			<td>1x Insulin Transferrin Selenium (ITS)</td>
			<td>Gibco</td>
			<td>51500056</td>
			<td></td>
		</tr>
		<tr>
			<td>1% KnockOut (KO) Serum Replacement</td>
			<td>Gibco</td>
			<td>10828-028</td>
			<td>Add fresh right before use.</td>
		</tr>
		<tr>
			<td>100% Dimethyl sulfoxide (DMSO)</td>
			<td>Sigma</td>
			<td>276855-100ML</td>
			<td></td>
		</tr>
		<tr>
			<td>CellTiter-Glo</td>
			<td>Promega</td>
			<td>G7573</td>
			<td>Purchased as a prepared solution.</td>
		</tr>
		<tr>
			<td>Consumables</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Small Combi Cassette</td>
			<td>ThermoFisher</td>
			<td>24073295</td>
			<td></td>
		</tr>
		<tr>
			<td>Small Multiflow Cassette</td>
			<td>BioTek</td>
			<td>294085</td>
			<td></td>
		</tr>
		<tr>
			<td>1536-well sterile TC plates (white)</td>
			<td>Corning</td>
			<td>3727</td>
			<td></td>
		</tr>
		<tr>
			<td>1536-well sterile TC plates (black)</td>
			<td>Corning</td>
			<td>3893</td>
			<td></td>
		</tr>
		<tr>
			<td>Scivax Plates</td>
			<td>MBL International</td>
			<td>NCP-LH384-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Equipment</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Adhesives seals</td>
			<td>ThermoFisher</td>
			<td>AB0718</td>
			<td></td>
		</tr>
		<tr>
			<td>Spinning Incubator</td>
			<td>LiCONiC</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Stainless Steel Lids</td>
			<td>The Genomics Institute of Novartis Research Foundation (GNF)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>ATS Acoustic Dispensor</td>
			<td>EDS Biosystems</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Echo Acoustic Dispensor</td>
			<td>Labcyte</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Luminometer</td>
			<td>Envision-Perkin Elmer</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Peristaltic Pump- Multidrop Combi</td>
			<td>ThermoFisher</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Liquid Handler-Multiflow FX</td>
			<td>BioTek</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

generation of high-throughput three-dimensional tumor spheroids for drug screening

Kræftceller har rutinemæssigt blevet kulturperler i to dimensioner (2D) på en plastik overflade. Denne teknik, men mangler den sande miljø en tumor masse er udsat for in vivo. Solide tumorer vokser ikke som et ark fastgjort til plastik, men i stedet som en samling af klonede celler i en tre-dimensionelle (3D) plads interagere med deres naboer, og med forskellige rumlige egenskaber, såsom forstyrrelser af normal cellulære polaritet. Disse interaktioner forårsage 3D-kulturperler celler til at erhverve morfologiske og cellulære karakteristika, som er mere relevante for i vivo tumorer. Derudover en tumor masse er i direkte kontakt med andre celletyper såsom stromale og immun celler samt den ekstracellulære matrix fra alle andre celletyper. Matrixen deponeret består af makromolekyler såsom kollagen og fibronektin.
I et forsøg på at øge oversættelse af forskningsresultater i onkologi fra bænken til bedside, er mange grupper begyndt at undersøge brugen af 3D-modelsystemer i deres stof udviklingsstrategier. Disse systemer er anset for at være mere fysiologisk relevante, fordi de forsøger at sammenfatte de komplekse og heterogene miljø af en tumor. Disse systemer kan dog være ret kompliceret, og selv om imødekommenhed over for vækst i 96-brønd formater, og nogle nu selv i 384, de tilbyder få valgmuligheder for omfattende vækst og screening. Dette observeret gap har ført til udviklingen af de metoder, der beskrives her i detaljer at kultur tumor spheroids i en høj overførselshastighed kapacitet i 1536-godt plader. Disse metoder udgør et kompromis for de yderst kompleks matrix-baserede systemer, som er vanskelige at skærmen og konventionel 2D assays. En lang række kræft cellelinjer husly forskellige genetiske mutationer er med held screenet, undersøge sammensatte effektivitet ved hjælp af en kurateret bibliotek af forbindelser rettet mod Mitogen-Activated Protein Kinase eller MAPK pathway. Klumpformet kultur svar er derefter sammenlignet med svar af celler dyrkes i 2D, og differential aktiviteter er rapporteret. Disse metoder giver en unik protokol for afprøvning sammensatte aktivitet i høj overførselshastighed 3D omgivelser.

En række etablerede kræftcelle linjer kendt for at vokse godt på 2D kultur betingelser blev testet ved hjælp af de metoder, der er skitseret her. Repræsentative billeder fra en bred vifte af MAPK mutant kræft cellelinjer (Calu-6:KRAS, NCI-H1299:NRAS, SK-MEL-30:NRAS og KNS-62:HRAS) ses i figur 1. Disse billeder viser, at selv om cellelinjer har forskellige morfologier, hver dannet 3D strukturer i 1536-godt assay plader. Figur 2</stro...

Watch this Scientific Journal Video about Generation of High-Throughput Three-Dimensional Tumor Spheroids for Drug Screening at JoVE.com

Generation of High-Throughput Three-Dimensional Tumor Spheroids for Drug Screening

På tværs af en bred vifte af tegn på sygdom, mere fysiologisk relevante modeller bliver udviklet og implementeret i drug discovery programmer. Den nye modelsystem beskrevet her demonstrerer hvordan tredimensionale tumor spheroids kan kulturperler og screenes i en høj overførselshastighed 1536-godt plade-baseret system til at søge efter nye onkologiske lægemidler.

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En integreret Platform for Genome-wide kortlægning af kromatin stater ved hjælp af høj overførselshastighed ChIP-sekventering i Tumor væv

Histone modifications constitute a major component of the epigenome and play important regulatory roles in determining the transcriptional status of ...

A Three-dimensional Thymic Culture System to Generate Murine Induced Pluripotent Stem Cell-derived Tumor Antigen-specific Thymic Emigrants

En tredimensionel Thymisk kultur system til at generere murine induceret pluripotente stamcelle-afledte tumor antigen-specifikke Thymisk emigranter

The inheritance of pre-rearranged T cell receptors (TCRs) and their epigenetic rejuvenation make induced pluripotent stem cell (iPSC)-derived T cells a ...

Longitudinal Morphological and Physiological Monitoring of Three-dimensional Tumor Spheroids Using Optical Coherence Tomography

Langsgående morfologiske og fysiologiske overvågning af tre-dimensionelle Tumor Spheroids bruge optisk kohærens tomografi

Tumor spheroids have been developed as a three-dimensional (3D) cell culture model in cancer research and anti-cancer drug discovery. However, currently, ...

Three-Dimensional Bone Extracellular Matrix Model for Osteosarcoma

Tre-dimensionelle knogle ekstracellulære Matrix Model for osteosarkom

Osteosarcoma (OS) is the most common and a highly aggressive primary bone tumor. It is characterized with anatomic and histologic variations along with ...

Fully Human Tumor-based Matrix in Three-dimensional Spheroid Invasion Assay

Fuldt humant tumor-baserede matrix i tredimensionale Spheroid invasion assay

Two-dimensional cell culture-based assays are commonly used in in vitro cancer research. However, they lack several basic elements that form the tumor ...

Physiologic Patient Derived 3D Spheroids for Anti-neoplastic Drug Screening to Target Cancer Stem Cells

Fysiologisk patient afledte 3D Sfæroider til anti-neoplastisk Drug screening for at målrette kræft stamceller

In this protocol, we outline the procedure for generation of tumor spheroids within 384-well hanging droplets to allow for high-throughput screening of ...

Establishment and Characterization of Small Bowel Neuroendocrine Tumor Spheroids

Etablering og karakterisering af små tarm neuroendokrine tumor Sfæroider

Small bowel neuroendocrine tumors (SBNETs) are rare cancers originating from enterochromaffin cells of the gut. Research in this field has been limited ...

Drug Screening of Primary Patient Derived Tumor Xenografts in Zebrafish

Drug Screening af primære patient afledt tumor xenografts i Zebrafish

Patient derived xenograft models are critical in defining how different cancers respond to drug treatment in an in vivo system. Mouse models are the ...

A Three-dimensional Model of Spheroids to Study Colon Cancer Stem Cells

En tre-dimensionel model af sfæroider til undersøgelse tyktarmskræft stamceller

Colorectal cancers are characterized by heterogeneity and a hierarchical organization comprising a population of cancer stem cells (CSCs) responsible for ...

High-Throughput In Vitro Assay using Patient-Derived Tumor Organoids

High-Throughput In Vitro Assay using Patient-Derived Tumor Organoids

High-Throughput In Vitro Assay ved hjælp af patient-afledte tumor organoider

Patient-derived tumor organoids (PDOs) are expected to be a preclinical cancer model with better reproducibility of disease than traditional cell culture ...